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PubMed1996

Patterns of translational regulation in the mammalian testis.

The translational activity of more than 40 different mRNAs in rodent testes has been analyzed by determining the proportions of inactive free-mRNPs and active polysomal mRNAs in sucrose gradients. These mRNAs can be sorted into several groups comprising mRNAs with similar patterns of translational activity in particular cell types. mRNAs in testicular somatic cells sediment primarily with polysomes, indicating that they are translated efficiently, whereas the vast majority of mRNAs in late meiotic and haploid spermatogenic cells display high levels of free-mRNAPs, indicative of a block to the initiation of translation. Protamine mRNAs exemplify a group of mRNAs that is transcribed in round spermatids, stored as free-mRNPs for several days, and translated in elongated spermatids after the cessation of transcription. The extent to which the free-mRNPs in primary spermatocytes and round spermatids are due to developmental changes in translational activity is unclear. mRNAs at these stages can often be detected earlier than the corresponding protein, implicating either a delay in translational activation or difficulties in detecting the protein. In contrast, sucrose gradients consistently indicate little difference in the proportions of various mRNAs in free-mRNPs in primary spermatocytes and round spermatids, implying that the proportions of translationally active mRNAs remain essentially constant. Since the levels of some mRNAs appear to greatly exceed the amount that is translated, the biological significance of some free-mRNPs in meiotic and early haploid cells in unclear. There are numerous examples of controls over the translation of individual mRNAs in meiotic and haploid cells; the proportions of various mRNAs in free-mRNPs range from virtually none to virtually all, and individual mRNAs are activated at specific stages in elongated spermatids. Existing evidence is contradictory whether the mRNAs in the protamine/transition protein gene family are repressed by mRNP proteins of sequestration.

Animals↗

PubMed2003

Novel CNBP- and La-based translation control systems for mammalian cells.

Throughout the development of Xenopus, production of ribosomal proteins (rp) is regulated at the translational level. Translation control is mediated by a terminal oligopyrimidine element (TOP) present in the 5' untranslated region (UTR) of rp-encoding mRNAs. TOP elements adopt a specific secondary structure that prevents ribosome-binding and translation-initiation of rp-encoding mRNAs. However, binding of CNBP (cellular nucleic acid binding protein) or La proteins to the TOP hairpin structure abolishes the TOP-mediated transcription block and induces rp production. Based on the specific CNBP-TOP/La-TOP interactions we have designed a translation control system (TCS) for conditional as well as adjustable translation of desired transgene mRNAs in mammalian cells. The generic TCS configuration consists of a plasmid encoding CNBP or La under control of the tetracycline-responsive expression system (TET(OFF)) and a target expression vector containing a TOP module between a constitutive P(SV40) promoter and the human model product gene SEAP (human secreted alkaline phosphatase) (P(SV40)-TOP-SEAP-pA). The TCS technology showed excellent SEAP regulation profiles in transgenic Chinese hamster ovary (CHO) cells. Alternatively to CNBP and La, TOP-mediated translation control can also be adjusted by artificial phosphorothioate anti-TOP oligodeoxynucleotides. Confocal laser-scanning microscopy demonstrated cellular uptake of FITC-labeled oligodeoxynucleotides and their localization in perinuclear organelles within 24 hours. Besides their TOP-based translation-controlling capacity, CNBP and La were also shown to increase cap-independent translation from polioviral internal ribosomal entry sites (IRES) and La alone to boost cap-dependent translation initiation. CNBP and La exemplify for the first time the potential of RNA-binding proteins to exert translation control of desired transgenes and to increase heterologous protein production in mammalian cells. We expect both of these assets to advance current gene therapy and biopharmaceutical manufacturing strategies.

Animals↗

PubMed2001

Specific secondary structures in the capsid-coding region of giardiavirus transcript are required for its translation in Giardia lamblia.

Enhanced translation of giardiavirus (GLV)-luciferase chimeric mRNA in Giardia lamblia requires the presence of the initial 264 nucleotides of the viral capsid-coding region. A 13 nt downstream box (DB) sequence within this region, complementary to a 15 nt sequence near the 3' end of G. lamblia 16 S-like ribosomal RNA (rRNA), was found to be essential for the enhanced translation. However, DB is located 64-78 nt downstream of the initiation codon, whereas an exponential increase of translation efficiency depends on a further increment of the coding region from nucleotides 111 to 264. Thus, there could be additional structural requirements for translation enhancement in the region downstream from DB. Four major stem-loop structures, designated I to IV, were identified in the MFOLD-predicted secondary structure of the 264 nt capsid-coding region with an estimated minimum free energy (DeltaG degrees ) of -77.16 kcal x mol(-1). Our chemical probing analysis of the free 264 nt RNA molecule in solution supports the predicted presence of stem-loops I, II and III, but casts doubts on stem-loop IV. It suggests, instead, the presence of a stem-loop IVA at a nearby location in the molecule. Site-directed mutagenesis designed to disrupt stem-loop structures I, II, III or IVA resulted in drastic reduction of translation efficiency, which was restored by compensatory sequence changes to regenerate individual stem-loop structures. Mutations disrupting the originally designated stem-loop IV did not exert any detectable effect on translation. However, alterations of the sequence UCUCC between nucleotides 216 and 220 in the flexible loop region of the revised secondary structure led to a precipitous drop in translation. Another stem-loop predicted by MFOLD that consists of a major portion of the DB sequence was examined by chemical probing but found little experimental support. Changes of the DB sequence without affecting the postulated stem structure led to drastic losses of translation efficiency. Thus, a simple structural basis for the enhanced translation could be that stem-loops I, II, III and IVA and the UCUCC sequence may facilitate the interaction between DB and the anti-DB in 16 S-like rRNA in initiating translation of GLV mRNA in G. lamblia.

Aldehydes↗

PubMed1994

Coronavirus translational regulation: leader affects mRNA efficiency.

Cells infected with the murine coronavirus, mouse hepatitis virus (MHV), show decreased host protein synthesis concomitant with an increase in viral protein synthesis. We examined the in vitro translation property of the conserved MHV 5'-leader RNA sequence by constructing chimeric mRNAs in which the 72-nt 5'-leader of M protein mRNA (A59 strain) was positioned upstream of the human alpha-globin coding region in a T7 expression vector. Synthetic 5'-capped transcripts of these mRNA constructs were translated in cell-free extracts prepared from uninfected and MHV-infected murine DBT cells. Nonviral mRNAs translated readily in both uninfected and infected cell-free extracts. By contrast, replacement of the human alpha-globin 5'-untranslated region (UR) with the MHV 5'-leader increased translation ca. three- to fourfold in cell-free extracts from MHV-infected cells versus translation in extracts from uninfected cells. Chimeric globin mRNA containing the reverse complementary sequence of the viral leader RNA in the 5'-UR showed no such increase in translation, indicating sequence specificity for the effect. A 13-nt region (-UCUAAUCCAAACA-) immediately proximal to the start codon was found to be important for the increased translation of the MHV leader-containing mRNAs. These data indicate that the apparent down-regulation of host translation is not primarily due to an inhibition of host translation but also involves a significant stimulation of viral translation in cis by a structural feature of the MHV 5'-leader RNA sequence in conjunction with a virus-specified or virus-induced factor.

Animals↗

PubMed1999

Hepatitis A virus translation is rate-limiting for virus replication in MRC-5 cells.

Translation of hepatitis A virus (HAV) RNA is controlled by an internal ribosome entry site (IRES) located within the 5' untranslated region (UTR). In some cell types, the characteristically slow growth of HAV may be due to inefficient viral translation. We investigated whether this is true in MRC-5 cells, which are used for vaccine production. We measured the impact of two clusters of mutations in the 5' UTR on virus translation and replication: the AG group was selected during passage in African green monkey kidney cells, and the MR group was selected during subsequent passage in MRC-5 cells. The efficiency of cap-independent translation was assessed by inserting cDNA encoding an HAV IRES upstream of the chloramphenicol acetyl transferase gene and transcription was driven in vivo by a hybrid T7/vaccinia virus system. A luciferase gene was inserted upstream of the IRES to serve as an internal control. Each HAV UTR was also inserted into an infectious cDNA clone; the average rate of viral RNA accumulation was determined for each mutant virus. In MRC-5 cells, the rate of virus replication was highly correlated with the efficiency of cap-independent translation (P = 0.006). The MR but not the AG mutations significantly increased both translation and viral RNA accumulation. Reversion of just one MR mutation (687 G to A) eliminated all of the replication-stimulating and translation-enhancing effects of the MR mutations. In the control BS-C-1 cells, there was no discernible correlation between the rate of virus replication and the efficiency of cap-independent translation (P = 0.136): the AG and MR groups combined had a small impact on translation, but no detectable impact on virus replication. We conclude that in MRC-5 cells viral translation is rate-limiting for HAV replication.

Animals↗

PubMed1999

Modulation of translational efficiency by contextual nucleotides flanking a baculovirus initiator AUG codon.

In a previous study of translational regulation of a baculovirus gene, we observed that translation initiated at an unexpectedly high efficiency from an AUG codon found in what was believed to be a poor context (M.-J. Chang and G. W. Blissard, 1997, J. Virol. 71, 7448-7460). In the current study, we examined the roles of nucleotides flanking a baculovirus AUG initiator codon in modulating translation initiation in lepidopteran insect cells. The roles of nucleotides flanking the AcMNPV gp64 initiator codon were examined by site-directed mutagenesis and functional assays in transfected Sf9 cells. To eliminate potential cis-acting sequences and effects, the gp64 initiator context was cloned in-frame with a chloramphenicol acetyl transferase reporter gene and under the control of a heterologous promoter. All possible single-nucleotide substitutions were generated in positions -6 to -1 and +4 to +6, relative to the A of the initiator AUG codon, which was designated +1. Constructs were transfected into lepidopteran cells and translation products were quantified by an enzyme-linked immunosorbent assay procedure. Substitutions of pyrimidines or other nucleotides at the -3 position resulted in little or no detectable effect on translation efficiency. In contrast, specific substitutions at the +4 and +5 positions resulted in approximately 2- to 3-fold increases in translation. Substitution of A in the +4 position resulted in an approximately 3-fold increase in translation, and substitution of any nucleotide for T in the +5 position resulted in approximately 1.9- to 2.8-fold increases. Substitutions at other positions (-6 to -1 and +6) resulted in no detectable increase or decrease in translation efficiency. These experimental results suggest an optimal initiator context of 5'-N N N N N N A U G A a/c/g N-3' for efficient translation initiation in lepidopteran cells. Consensus translation initiation contexts were generated from baculovirus genes and lepidopteran genes, then compared with the experimental results from the gp64 initiator context.

Animals↗

PubMed1994

Analysis of hepatitis A virus translation in a T7 polymerase-expressing cell line.

Hepatitis A virus (HAV) exhibits several characteristics which distinguish it from other picornaviruses, including slow growth in cell culture even after adaptation, and lack of host-cell protein synthesis shut-down. Like other picornaviruses, HAV contains a long 5' nontranslated region (NTR) incorporating an internal ribosomal entry site (IRES), which directs cap-independent translation. We compared HAV IRES-initiated translation with translation initiated by the structurally similar encephalomyocarditis virus (EMCV) IRES, using plasmids in which each of the 5'NTRs is linked in-frame with the chloramphenicol acetyltransferase (CAT) gene. Translation was assessed in an HAV-permissive cell line which constitutively expresses T7 RNA polymerase and transcribes high levels of uncapped RNA from these plasmids following transfection. RNAs containing the EMCV IRES were efficiently translated in these cells, while those containing the HAV IRES were translated very poorly. Analysis of translation of these RNAs in the presence of poliovirus protein 2A, which shuts down cap-dependent translation, demonstrated that their translation was cap independent. Our results suggest that the HAV IRES may function poorly in these cells, and that inefficient translation may contribute to the exceptionally slow replication cycle characteristic of cell culture-adapted HAV.

Animals↗

PubMed2005

Orienting eye movements and nystagmus produced by translation while rotating (TWR).

Sinusoidal translation while rotating at constant angular velocity about a vertical axis (translation while rotating, TWR) produces centripetal and translational accelerations along the direction of translation and an orthogonal Coriolis acceleration due to the translation in the rotating frame. Thus, a Coriolis acceleration is produced along the bitemporal axis when oscillating along the naso-occipital axis, and along the naso-occipital axis when oscillating along the bitemporal axis. Together, these components generate an elliptically rotating acceleration vector that revolves around the head in the direction of rotation at the frequency of oscillation. Here we studied the orienting and compensatory responses of rabbits during TWR. Combinations of centripetal and translational accelerations were held constant at 0.5 g, and oscillation frequencies were varied from 0.01-0.33 Hz. The amplitude of the Coriolis acceleration increased with the frequency of translation. Naso-occipital translation caused vergence and pitch at all frequencies and roll at higher frequencies, and bitemporal translation produced roll at all frequencies and vergence and pitch at higher frequencies. The sensitivity of each ocular orienting component to linear acceleration was comparable across the different oscillation frequencies. TWR also induced continuous yaw nystagmus with slow phase velocity in the direction of rotation of the acceleration vector. Thresholds for appearance of nystagmus were 0.05 Hz, corresponding to a Coriolis acceleration of 0.06 g. Mean slow phase velocity for a rotating linear acceleration vector produced by 0.5 g along the translation axis and 0.34 g of Coriolis acceleration along the orthogonal axis were approximately 9 degrees /s. Eye velocities during TWR were similar to those generated by off-vertical axis rotation (OVAR), but were opposite in direction with regard to head rotation, following the direction of the rotating acceleration vector in both paradigms. Both are produced by activation of velocity storage in the vestibular system. One important difference between TWR and OVAR is that the head is always upright with regard to gravity during TWR. We speculate that the brain may use these low amplitude rotating linear accelerations to generate eye velocities that help to orient gaze when making turns during normal locomotion.

Acceleration↗

PubMed2005

Evaluation of cis-acting elements in the rubella virus subgenomic RNA that play a role in its translation.

The subgenomic (SG) mRNA of rubella virus (RUB) contains the structural protein open reading frame (SP-ORF) that is translated to produce the three virion structural proteins: capsid (C) and glycoproteins E2 and E1. RUB expression vectors have been developed that express heterologous genes from the SG RNA, including replicons which replace the SP-ORF with a heterologous gene, and these expression vectors are candidate vaccine vectors. In the related alphaviruses, translational enhancing elements have been identified in both the 5' untranslated region (UTR) of the SG RNA and the N-terminal region of the C gene. To optimize expression from RUB vectors, both the 5'UTR of the SG RNA and the C gene were surveyed for translational enhancing elements using both plasmids and replicons expressing reporter genes from the SG RNA. In replicons, the entire 5'UTR was necessary for translation; interestingly, when plasmids were used the 5'UTR was dispensable for optimal translation. The RUB C gene contains a predicted long stem-loop starting 62 nts downstream from the initiation codon (SLL) that has a structure and stability similar to SL's found in the C genes of two alphaviruses, Sindbis virus (SIN) and Semliki Forest virus, that have been shown to enhance translation of the SG RNA in infected cells. However, a series of fusions of various lengths of the N-terminus of the RUB C protein with reporter genes showed that the SLL had an attenuating effect on translation that was overcome by mutagenesis that destabilized the SLL or by adding downstream sequences of the C gene to the fusion. Thus, for optimal expression efficiency from RUB expression vectors, only the 5'UTR of the SG RNA is required. Further investigation of the differing effects of the SLL on RUB and alphavirus SG RNA translation revealed that the SIN and RUB SLLs could enhance translation when expressed from a SIN cytopathic replicon, but not when expressed from a plasmid, a RUB replicon, or a SIN noncytopathic replicon. Thus, the SLL only functions in a "cytopathic environment" in which cell translation has been altered.

Animals↗

PubMed1990

Polyadenylated RNA sequences from vaccinia virus-infected cells selectively inhibit translation in a cell-free system: structural properties and mechanism of inhibition.

The mechanism of vaccinia virus-induced selective inhibition of host cell protein synthesis was studied in a nonpermissive (Chinese hamster ovary, CHO) and in a permissive mouse cell line ( L cells). Small polyadenylated RNAs obtained from uninfected and infected cells were fractionated into six size classes by polyacrylamide gel electrophoresis. The RNAs from the first two largest fractions (greater than 500 nucleotide, nt) were translated into some low-molecular-weight polypeptides, whereas, the RNAs from the remaining fractions (400-500, 300-400, 200-300, and 100-200 nt) had no translational activity in reticulocyte lysates. When these nontranslating polyadenylated short sequences (POLADS) were added to the cell-free system together with HeLa cell mRNAs, translation was inhibited from 70%, by the 400- to 500-nt fraction, to about 20%, by the 100- to 200-nt fraction. The degree of inhibition of protein synthesis was clearly dependent on the size of POLADS. The translation of vaccinia virus mRNAs in the cell-free system was inhibited by about 25% with the 400- to 500-nt fraction, by 5% with the 300- to 400-nt fraction, while the smaller size POLADS had no inhibitory effect. The inhibition of HeLa cell and vaccinia virus mRNA translation by POLADS was reversed by the simultaneous addition of oligo(dT) to the cell-free system. POLADS were also obtained from uninfected cells, but they inhibited the translation of HeLa cell and vaccinia virus mRNAs to a much lesser extent. The removal of the poly(A) moiety from POLADS by treatment with ribonuclease H and oligo(dT) abolished their inhibitory effect on HeLa cell mRNA translation. The average length of the poly(A) tails of POLADS obtained from infected cells was longer than that of POLADS from normal cells. Inhibition of HeLa cell mRNA translation mediated by POLADS in the cell-free system was reversed (approximately 70%) by addition of crude initiation factors (ribosomal salt wash, RSW). Significantly, inhibition of translation of POLADS was reversed (greater than 90%) by addition of purified poly(A) binding protein (PAB). Purified initiation factor 4A (eIF-4A) also reversed this inhibition, but to a lesser extent than RSW and PAB. Our results show that the translation of vaccinia virus mRNAs is resistant to POLADS, suggesting that POLADS, by virtue of their long poly(A) tails, may sequester PAB and thus, play a role in selective inhibition.

Animals↗

PubMed1986

Translocation of murine interleukin 2 into microsomes during translation in a cell-free system.

Interleukin 2 (IL2) is a lymphokine which stimulates the growth of T lymphocytes. Although IL2 mRNA is translated into biologically active IL2 relatively efficiently in microinjected Xenopus laevis oocytes, it has been difficult to establish reproducible cell-free translation systems for this lymphokine. Such systems would be useful for the analysis of translational and post-translational events. In this paper, we show that a wheat germ extract will translate IL2 mRNA into biologically active murine IL2, most of which is translocated into dog pancreas microsomes when these are present. Translocation occurs only during translation, but wheat germ extracts translate IL2 mRNA whether microsomes are present or not. Surprisingly, reticulocyte lysates do not readily translate IL2 mRNA, and strongly inhibit its translation in wheat germ extracts. This inhibition can be partially alleviated by adding dog pancreas microsomes to the system. The inhibition seen with reticulocyte lysate may be attributable to the action of the signal recognition particle, which binds nascent secretory proteins, and blocks their further translation in the absence of the docking protein present in microsomes.

Animals↗

PubMed2004

Targeting internal ribosome entry site (IRES)-mediated translation to block hepatitis C and other RNA viruses.

A number of RNA-containing viruses such as hepatitis C (HCV) and poliovirus (PV) that infect human beings and cause serious diseases use a common mechanism for synthesis of viral proteins, termed internal ribosome entry site (IRES)-mediated translation. This mode of translation initiation involves entry of 40S ribosome internally to the 5' untranslated region (UTR) of viral RNA. Cap-dependent translation of cellular mRNAs, on the other hand, requires recognition of mRNA 5' cap by the translation machinery. In this review, we discuss two inhibitors that specifically inhibit viral IRES-mediated translation without interfering with cellular cap-dependent translation. We present evidence, which suggest that one of these inhibitors, a small RNA (called IRNA) originally isolated from the yeast Saccharomyces cerevisiae, inhibits viral IRES-mediated translation by sequestering both noncanonical transacting factors and canonical initiation factors required for IRES-mediated translation. The other inhibitor, a small peptide from the lupus autoantigen La (called LAP), appears to block binding of cellular transacting factors to viral IRES elements. These results suggest that it might be possible to target viral IRES-mediated translation for future development of therapeutic agents effective against a number of RNA viruses including HCV that exclusively use cap-independent translation for synthesis of viral proteins.

5' Untranslated Regions↗

PubMed2006

Gender differences in post-exercise sagittal knee translation: a comparison between elite volleyball players and swimmers.

BACKGROUND: There is an increased risk for anterior cruciate ligament injury during the last part of a match or training session and one reason for that could be a post-exercise increase in tibial translation. PURPOSE: To investigate if sagittal tibial translation is affected after a workout session in volleyball or swimming in elite athletes. In addition, gender differences in sagittal tibial translation after the workout session were investigated. METHOD: Thirty-one elite volleyball players (16 male) and 33 elite swimmers (15 male) participated in this study. Measurements of total tibial translation were taken before and after a workout session in either volleyball or swimming with the use of a KT-1000 arthrometer. RESULTS: Total tibial translation increased by 1.1 mm (SD 1.9) in the group consisting of both male and female volleyball players (p=0.003) and remained unchanged in the swimmers. Male athletes increased their tibial translation with 1.8 mm (SD 1.8) and 0.6 mm (SD 1.1) in the two sports, respectively, while the tibial translation did not increase in the female athletes. CONCLUSION: Impact sports such as volleyball training leads to a post-exercise increase in tibial translation in male athletes. The increase in tibial translation in swimmers, that is a non-impact sport, was small and may not be clinically significant for the functional stability of the joint. It has been shown that female athletes have an increased risk for injury. Our results show no support for an increase in tibial translation being an important factor for this increased risk, and suggest that the difference between males and females in this regard should be sought elsewhere.

Adolescent↗

PubMed2005

The hypoxic proteome is influenced by gene-specific changes in mRNA translation.

BACKGROUND AND PURPOSE: Hypoxia causes a rapid reduction in mRNA translation efficiency. This inhibition does not affect all mRNA species to the same extent and can therefore contribute significantly to hypoxia-induced differential protein expression. Our aim in this study was to characterize changes in gene expression during acute hypoxia and evaluate the contribution of regulation via mRNA translation on these changes. For each gene, the contribution of changes in mRNA abundance versus mRNA translation was determined. MATERIALS AND METHODS: DU145 prostate carcinoma cells were exposed to 4h of hypoxia (<0.02% O2). Efficiently translated mRNAs were isolated by sedimentation through a sucrose gradient. Affymetrix microarray technology was used to evaluate both the transcriptional and translational contribution to gene expression. Results were validated by quantitative PCR. RESULTS: One hundred and twenty genes were more than 4-fold upregulated by hypoxia in the efficiently translated fraction of mRNA, in comparison to only 76 genes at the level of transcription. Of the 50 genes demonstrating the largest changes in translation, 11 were found to be more than 2-fold over represented in the translated fraction in comparison to their overall transcriptional level. The gene with the highest translational contribution to its induction was CITED-2, which is a negative regulator of HIF-1 transcriptional activity. CONCLUSIONS: Gene-specific regulation of mRNA translation contributes significantly to differential gene expression during hypoxia.

Cell Hypoxia↗

PubMed1996

Mutations in yeast ribosomal proteins S28 and S4 affect the accuracy of translation and alter the sensitivity of the ribosomes to paromomycin.

Ribosomal proteins S12, S5 and S4 of Escherichia coli are essential for the control of translational accuracy. Their yeast equivalents, i.e., S28, S4 and S13, have also been implicated in this process. Using a poly(U)-dependent cell-free translation system, we determined the accuracy of translation and the sensitivity to antibiotic paromomycin of yeast ribosomes carrying mutant ribosomal proteins S28 and/or S4. Our results confirm by quantitative biochemical methods previous genetic data showing that proteins S28 and S4 are involved in the decoding activity of the ribosome and interact to control translational accuracy. We find that the suppressor mutation SUP44 in yeast S4, decreased the accuracy of translation. To examine the effect of mutant S28, we disrupted RPS28B and introduced in RPS28A the same substitutions that cause hyperaccurate translation or antibiotic resistance in bacteria. Three of these substitutions (Lys-62-->Asn, Thr or Gln) similarly increased translational accuracy in vitro or antibiotic resistance. In the presence of the SUP44 mutation, these substitutions partially reversed the decrease of translational accuracy caused by SUP44. However, the Lys-62-->Arg substitution decreased translational accuracy and caused antibiotic sensitivity both in nonsuppressor and in SUP44 haploids. These results establish the role of Lys-62 of S28 in optimizing translational accuracy and provide a more precise view of the functional role of two important ribosomal proteins.

Anti-Bacterial Agents↗

PubMed2000

The effect of negative intraarticular pressure and rotator cuff force on glenohumeral translation during simulated active elevation.

OBJECTIVE: The objective of this study was to determine the effect of negative pressure in the intraarticular space and subacromial bursa, as well as rotator cuff force, on glenohumeral translation during active elevation. DESIGN: Glenohumeral translation during elevation in the scapular plane was measured under greater than, less than, and equal to physiologic rotator cuff muscle force, as well as with and without the subacromial bursa and intraarticular space vented. BACKGROUND: Negative intraarticular pressure has been shown to help stabilize the glenohumeral joint in passive motion, although the effect on translation during active motion has not been investigated. METHODS: Eight cadaverous human shoulder specimens were tested in a dynamic shoulder simulator. Forces in the muscles of the rotator as well as the middle deltoid muscle were simulated using servohydraulic cylinders. Joint motion was measured using an ultrasonic motion analysis system. RESULTS: Superior translation of the humerus increased 1.2 mm (SD, 0.4) upon venting of the bursa, and 2.1 mm (SD, 1.7) upon venting of the joint capsule at 25 degrees of glenohumeral elevation in the scapular plane. At 90 degrees elevation, venting the bursa did not change superior translation but increased anterior translation 1.04 mm (SD, 1.0). Venting of the joint capsule increased superior translation by 2.8 mm (SD, 2.5). Decreasing rotator cuff force increased superior translation, while reducing it did not. CONCLUSION: The muscles of the rotator cuff and the negative pressure of the intraarticular space and the subacromial bursa stabilize the glenohumeral joint since they restrict translation in the superior and anterior directions. RelevanceIncreased glenohumeral translation and the resulting asymmetric loading may lead to arthrosis and ultimately rotator cuff arthropathy.

Adult↗

PubMed2002

The progression of anterior translation after anterior cruciate ligament reconstruction in a caprine model.

Large post-operative anterior-posterior translations are frequently reported after quadruped anterior cruciate ligament (ACL) reconstructions. To determine when the translation increases occur and the mechanism responsible, we followed the anterior and posterior translation limits in 18 goat knees for six months. Reconstructions were performed using grafts 4 or 7 mm wide placed in initially tight or lax positions. The anterior and posterior translation limits at 50 N were monitored using Roentgen stereophotogrammetric analysis. Graft bone block stability and soft tissue segment lengths were also assessed. Large (> 2 mm) increases in anterior translation were noted in 71% of the subjects at two weeks, and in 88% at eight weeks. The translations in the lax and tight groups were indistinguishable after two weeks. Joints with wide grafts had less anterior translation compared to narrow grafts at all time periods, but were significant different only at 26 weeks. The posterior translation limit moved anteriorly over the 26 weeks. Eight of nine joints had stable graft bone markers and/or increases in graft soft tissue lengths. In conclusion, increased anterior translation occurred soon after ACL reconstruction, was associated with graft soft tissue changes, and appeared to be reduced by larger grafts. A post-surgical decrease in posterior translation limit was also observed.

Animals↗

PubMed1998

Features of the autonomous function of the translational enhancer domain of satellite tobacco necrosis virus.

The RNA of satellite tobacco necrosis virus (STNV) is a monocistronic messenger that lacks both a cap and a poly(A) tail. Translation of STNV RNA in vitro is promoted by a 120-nt translational enhancer domain (TED) in the 3'-untranslated region. TED also stimulates translation of heterologous mRNAs. In this study, we show that TED stimulates translation of a cat mRNA by increasing translation efficiency to the level of capped mRNA. This stimulatory activity is not impaired by translation through TED. TED stimulates translation efficiency from different positions within the mRNA, varying from the 5' end to 940 nt downstream of the coding region. Duplication of TED has an additive effect on translation stimulation only when located at both ends of the mRNA. On dicistronic RNAs, TED stimulates translation of both cistrons to the same extent. These data suggest that TED acts primarily by recruiting the translational machinery to the RNA.

3' Untranslated Regions↗