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PubMed1996

The RNA world of the nucleolus: two major families of small RNAs defined by different box elements with related functions.

We have discovered that all known yeast and vertebrate small nucleolar RNAs (snoRNAs), except for the MRP/7-2 RNA, fall into two major classes. One class is defined by conserved boxes C and D and the other by a novel element: a consensus ACA triplet positioned 3 nt before the 3' end of the RNA. A role for the ACA box is snoRNA stability has been established by mutational analysis of a yeast ACA snoRNA (snR 11). Full function of the box depends on the integrity of an adjacent upstream stem. All members of the yeast ACA family are associated with the GAR1 protein. Binding of this or another common small nucleolar ribonucleoprotein particle protein is predicted to be a critical entry point to snoRNA posttranscriptional life, including precise formation of the snoRNA 3' end.

Animals↗

PubMed2001

Two CCAAT boxes in a novel inverted repeat motif are required for Hlx homeobox gene expression.

Hlx is a homeobox transcription factor gene required for normal intestinal and hepatic growth in development. We previously found high sequence identity and 17 conserved consensus cis-regulatory/transcription factor binding elements in the mouse and human Hlx 5' regions. A 594 bp sequence in the Hlx 5' region possessing the same activity in driving luciferase expression as larger Hlx 5' sequences had three segments each necessary but not sufficient for luciferase expression in NIH 3T3 cells (which express Hlx). Nine of the conserved putative regulatory elements are positioned within these segments, including two CCAAT boxes on opposite strands within a conserved 44 bp inverted repeat sequence. To test the hypothesis that these elements are required for promoter activity, we compared the reporter expression activity of segments containing mutations of these elements with activity of the parent Hlx promoter sequence. We found that mutation of either CCAAT box or a conserved AP-2 site resulted in a significant decrease in promoter activity. Restoration of the inverted repeat with complementary mutations of both CCAAT boxes did not restore activity. Further, mutation of other portions of the inverted repeat did not affect promoter activity. Mutation of other elements had no effect on promoter activity.

Animals↗

PubMed1998

Cloning of a novel Y-box homology protein (chkYB-1HP) cDNA lacking the cold-shock domain.

We have isolated the complete cDNA clone of a novel 262 amino acid Chicken YB-1 Homology Protein (chkYB-1HP) by screening a chicken embryo cDNA expression library. While the chkYB-1HP is identical over its carboxyl-terminal 78 amino acids with the Y-box protein YB-1, it differs strikingly from all other Y-box transcription factors by lacking the cold-shock domain (CSD). We propose that proteins like chkYB-1HP that lack the CSD, but retain the hydrophilic carboxyl domain could regulate Y-box proteins through the formation of heterodimeric complexes.

Amino Acid Sequence↗

PubMed1998

The mouse Sox5 gene encodes a protein containing the leucine zipper and the Q box.

The nucleotide sequence of mouse Sox5 (mSox5) cDNA derived from the testis has been reported by Denny et al. (EMBO J. 11 (1992) 3705-3712). We newly isolated an mSox5 cDNA derived from 8.5-day mouse embryo. Our cDNA encodes a protein of 763 amino acids, which is considerably larger in size than the previous one (392 amino acids) derived from the adult mouse testis. The most significant difference between the embryo-derived and testis-derived mSox5 cDNAs is that the embryonic one encodes a leucine zipper motif and a neighboring glutamine-rich sequence stretch (named Q box), but the testis-derived one does not. The leucine zipper and the Q box are highly conserved among type-D SOX proteins including mSox5. mSox5 was suggested to be a single-copy gene by Southern analysis. With reverse transcription/polymerase chain reaction, we found that not only mouse embryo, but also the adult mouse testis, express mSox5 mRNA species encoding the leucine zipper and the Q box.

Amino Acid Sequence↗

PubMed2000

Characterization and transcriptional profiles of two rice MADS-box genes.

The plant MADS-box gene family plays a key role in plant development, especially in flower development. We designed degenerate primer according to the MADS-box conserved region and isolated two cDNA from rice, FDRMADS6 and FDRMADS7, which are homologous to AP1. RT-PCR expression analyses by using total RNA isolated from root, shoot and flower showed that the FDRMADS6 transcript was detectable only in flower while FDRMADS7 was expressed in all three tissues. In situ hybridization experiments indicated that at the early stage of rice flower development, the transcripts of FDRMADS6 and FDRMADS7 were detected in the spikelet apical meristem, which were same as AP1. At the late stage, when flower organ primordia started differentiating, the expression of FDRMADS6 appeared to be specifically localized in developing stamens and the pistil primordia, while the transcripts of FDRMADS7 were detectable abundantly throughout the organ primordia. Our results suggest the two MADS-box genes may be members of the AP1 family, but may have different functions.

Journal Article↗

PubMed1997

The last proline of Box 1 is essential for association with JAK2 and functional activation of the prolactin receptor.

The interaction of prolactin (PRL) with its receptor leads to activation of the tyrosine kinase, Janus kinase 2 (JAK2). In the cytoplasmic juxtamembrane region, a short segment (Box 1) which is conserved in other receptors of the PRL/growth hormone (GH)/cytokine receptor family, is required for signal transduction. To assess the contribution of the different amino acids of Box 1, individual alanine substitutions of all residues, grouped substitution of four prolines (4PA mutant) and individual leucine replacement of the two last prolines (P248L and P250L mutants) were introduced. Here we show that P250L and 4PA (i) inhibit PRL-induced transactivation of a luciferase reporter governed by a beta-caseine gene promoter; (ii) decrease in JAK2 tyrosine kinase activity in biotinylated-PRL precipitates; (iii) impair the interaction between PRLR and JAK2, as evidenced by lack of co-immunoprecipitation, (iv) and prevent the activation of signal transducer and activator of transcription (Stat) as determined by absence of tyrosine phosphorylation of Stat5. Our data suggest that the Box 1 region of the PRL receptor and particularly the last proline is critical for JAK2 association and subsequent activation. These results support the notion that the tyrosine kinase JAK2 is implicated in activation of downstream protein effectors such as Stat5, which are involved in transcription of PRL-responsive genes.

DNA-Binding Proteins↗

PubMed1995

Percutaneous Kirschner wire fixation through the snuff box: an anatomic study.

We undertook this study to better define the anatomy of the radial aspect of the wrist and to establish a zone of safety for the placement of Kirschner wires, cannulated screws, and arthroscopes within the anatomic snuff box. Twenty fresh frozen cadaver upper extremities underwent placement of three percutaneous Kirschner wires under fluoroscopic guidance through the anatomic snuff box. In each extremity, one Kirschner wire was placed into the radial styloid, one across the scaphocapitate joint, and one across the scapholunate joint. A safe zone of mean 0.68 sq cm was found deep to the subcutaneous tissue bordered by the radial styloid, the first dorsal compartment, the radial artery, and the superficial radial nerve. Kirschner wires placed distal, dorsal, or palmar to the borders of the safe zone were at great risk of injuring neurovascular structures. To minimize the risk of injury to adjacent structures, we advise a limited incision in the safe zone with blunt dissection to the wrist capsule. Though improved anatomic understanding, we established a new location for the arthroscopic 1,2 portal within the snuff box.

Arthroscopy↗

PubMed2002

A new subfamily of structurally related human F-box proteins.

F-box proteins, a critical component of the evolutionary conserved ubiquitin-protein ligase complex SCF (Skp1/Cdc53-Cullin1/F-box), recruit substrates for ubiquitination and consequent degradation through their specific protein-protein interaction domains. Here, we report the identification of full-length cDNAs encoding three novel human F-box proteins named FBG3, FBG4 and FBG5 which display similarity with previously identified NFB42 (FBX2) and FBG2 (FBX6) proteins. All five proteins are characterized by an approximately 180-amino-acid (aa) conserved C-terminal domain and thus constitute a third subfamily of mammalian F-box proteins. Analysis of genomic organization of the five FBG genes revealed that all of them consist of six exons and five introns. FBG1, FBG2 and FBG3 genes are located in tandem on chromosome 1p36, and FBG4 and FBG5 are mapped to chromosome 19q13. FBG genes are expressed in a limited number of human tissues including kidney, liver, brain and muscle tissues. Expression of rat FBG2 gene was found related to differentiation/proliferation status of hepatocytes. Specifically, FBG2 mRNA was expressed in foetal liver, decreased after birth and re-accumulated in adult liver. Expression of FBG2 was strongly inhibited in hepatoma cells by okadaic acid.

Amino Acid Sequence↗

PubMed2003

Role of MADS box proteins and their cofactors in combinatorial control of gene expression and cell development.

In all organisms, correct development, growth and function depends on the precise and integrated control of the expression of their genes. Often, gene regulation depends upon the cooperative binding of proteins to DNA and upon protein-protein interactions. Eukaryotes have widely exploited combinatorial strategies to create gene regulatory networks. MADS box proteins constitute the perfect example of cellular coordinators. These proteins belong to a large family of transcription factors present in most eukaryotic organisms and are involved in diverse and important biological functions. MADS box proteins are combinatorial transcription factors in that they often derive their regulatory specificity from other DNA binding or accessory factors. This review is aimed at analyzing how MADS box proteins combine with a variety of cofactors to achieve functional diversity.

Amino Acid Sequence↗

PubMed1993

Arthroscopy training using a "black box" technique.

In orthopaedic surgery residency, arthroscopic training is an essential component and one of the educational program requirements. A system using standard arthroscopy instrumentation and a "black box" has been developed for the purpose of teaching basic arthroscopic surgical skills in a laboratory setting. The black box component can be used as a dry or wet model, permitting a variety of arthroscopic techniques to be mastered under controlled circumstances. This unit has provided excellent feedback from residents and staff, and is cost effective. The basic principles of the orthopedic black box can be adapted for the development of basic skills in other specialties.

Arthroscopy↗

PubMed2004

Effects of bacterial superantigens on behavior of mice in the elevated plus maze and light-dark box.

Bacterial superantigens, such as staphylococcal enteroxins A and B (SEA/SEB) stimulate T cells to produce high levels of cytokines in blood. Previously it had been shown that these toxins were capable of stimulating increased neuroendocrine activity and enhanced behavioral reactivity to novel gustatory and non-gustatory stimuli. Therefore, it was suggested that these superantigens may promote anxiety-like behavior. In the current set of experiments, BALB/cByJ and C57BL/6J male mice were challenged with either SEB (50 microg) or SEA (5 or 10 microg) and tested for behavior in the elevated plus maze (EPM). Results suggested an absence of increased anxiety-like behavior, with exploration of the open arms being enhanced by SEA or SEB treatment. In another test of anxiety, the light-dark box, SEB challenge of BALB/cByJ mice 90 min prior to testing, did not alter exit latency, activity nor time spent in the dark. However, in a second experiment, it was found that if animals were first tested for consumption, followed by testing in the light-dark box, SEB challenged animals displayed increased exit latency and reduced exploration. These studies suggest that in standard tests of rodent anxiety-like behavior, evidence for the induction of anxiety-like processes subsequent to challenge with SEA or SEB is not patently discernable. However, neurobiological events induced by immunological challenge might synergize with reactivity to psychogenic and/or gustatory stimuli, thereby resulting in increased anxiety-like behavior that could be unmasked by standard behavioral tests such as the light-dark box or EPM.

Analysis of Variance↗

PubMedDate not supplied

Self-regulatory behaviors in children with Down syndrome and typically developing children measured using the Goodman Lock Box.

Self-regulation has been identified as an area of difficulty for those with mental retardation. The Goodman Lock Box provides measures of two critical aspects of self-regulation-planfulness and maintenance of goal-directed behavior. In this study, the Lock Box performance of 25 children with Down syndrome was compared with that of 43 typically developing children, matched for mental age (24-36 months). Children in both groups showed similar levels of competence, planfulness and distractibility. However, children with Down syndrome displayed more task-avoidant behavior. Some issues related to the measurements obtained from the Lock Box are raised.

Child Behavior↗

PubMed2000

Electronic pill-boxes in the evaluation of antihypertensive treatment compliance: comparison of once daily versus twice daily regimen.

The objective was to compare the compliance of hypertensive patients treated with captopril twice daily or trandolapril once daily. After a 2-week placebo period, hypertensive patients (diastolic BP 95-115 mm Hg) were randomly allocated to trandolapril 2 mg once daily or to captopril 25 mg twice daily for 6 months. Trandolapril and captopril were packed in electronic pill-boxes equipped with a microprocessor that recorded date and time of each opening (MEMS). Patients' compliance was assessed both by standard pill-count and by electronic monitoring. Blood pressure was measured using a validated semi-automatic device at the end of the placebo period and of the treatment period. One hundred sixty-two patients entered the study. Compliance data were evaluable for 133 patients (62 in the captopril group and 71 in the trandolapril group). Treatment groups were comparable at baseline except for age (P = .046). Using electronic pill-box, overall compliance was 98.9% in the trandolapril group and 97.5% in the captopril group (P = .002). The percentage of missed doses was 2.6% in the trandolapril group and 3.3% in the captopril group (P = .06). The percentage of delayed doses was 1.8% in the trandolapril group and 11.7% in the captopril group (P = .0001). The percentage of correct dosing periods, ie, a period with only one correct recorded opening, was 94.0% in the trandolapril group and 78.1% in the captopril group (P = .0001). Results were unchanged when adjusted for age. At the end of the study, 41% of patients in the trandolapril group and 27% in the captopril group (NS) had their blood pressure normalized (systolic BP <140 and diastolic BP <90 mm Hg). In this 6-month study, the electronic pill-box allowed refined analysis of compliance of hypertensive patients. Patients' compliance with once daily trandolapril was higher than with twice daily captopril. The between-group difference is mainly explained by an increase in delayed doses in the twice daily group.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed1997

Acute stress enhances anxiolytic-like drug responses of mice tested in a black and white test box.

Drug-induced facilitation of exploration of a two-compartment black and white test box was studied in mice applying different stressors before test. Tail suspension suppressed exploration of both compartments and enhanced diazepam- or eltoprazine-induced increase of exploration of the white compartment. (-)-Penbutolol reversed the effects of eltoprazine suggesting involvement of 5-HT(1A/1B) receptors. Buspirone had weak effects both under standard conditions, and after tail suspension. Ritanserin suppressed exploratory behavior, and this effect was potentiated by tail suspension. In conclusion, application of acute tail suspension stress immediately before test in the black and white box enhanced the sensitivity to drugs. In contrast to this, forced swimming suppressed general behavioral activity, but in this case diazepam facilitated the behavior in both compartments. Foot shocks immediately before test reduced activity in the black compartment and did not affect the behavior in the white compartment. Thus different acute stressors modified behavior in the black and white box differently. Chronic stress (crowded housing) produced an anxiolytic-like response.

Animals↗

PubMed1997

Effects of beta-adrenoceptor blocker pindolol, calcium antagonist verapamil and their combination on retention in step-down- and shuttle-box-trained rats and on brain biogenic monoamines.

The effects of the beta-adrenoceptor blocker pindolol and the calcium antagonist verapamil administered alone or in combination on retention in step-down- and shuttle-box-trained rats and on the biogenic monoamine levels in the frontal cortex and hippocampus were examined. The chronic oral treatment with pindolol impaired retention in step-down- and shuttle-box-trained rats, decreasing the dopamine (DA) and noradrenaline (NA) levels and increasing the serotonin (5-HT) levels in the cortex and hippocampus. Verapamil did not influence retention in step-down- and shuttle-box avoidance situation and the biogenic monoamine levels in the frontal cortex and hippocampus. It should, however, be noted that the chronic oral treatment with verapamil completely abolished the retention-impairing effect of pindolol, restoring to normal DA, NA and 5-HT levels. These findings might be of interest to clinical practice and suggest the necessity for using a combination of beta-blockers with Ca2+ antagonists in case of prolonged treatment of cardiovascular diseases.

Adrenergic beta-Antagonists↗

PubMed1998

The low level expression of chloramphenicol acetyltransferase (CAT) mRNA in Escherichia coli is not dependent on either Shine-Dalgarno or the downstream boxes in the CAT gene.

Recent studies have shown that the canonical Shine-Dalgarno (SD)-anti-SD interaction is dispensable for the initiation of translation of certain mRNAs in Escherichia coli. Alternative non-SD sequences (located upstream from the initiation codon) and also downstream sequences ("downstream boxes") complementary to 16S rRNA were found to be involved in the initiation of translation of mRNAs devoid of either SD or any leader sequences. In this study the chloramphenicol acetyltransferase (CAT) gene was modified to remove the 5' terminal non-translated region and/or the two potential downstream boxes in the CAT gene. Thus a series of ten CAT gene constructs was created and expressed in E. coli under a strong constitutive promoter. The results showed that CAT mRNAs devoid of both leader sequence nucleotides and the two downstream boxes in the CAT gene remained active in vivo and produced CAT protein in sufficient amounts for survival of the transformed cells at chloramphenicol concentrations up to 20-30 micrograms/ml.

5' Untranslated Regions↗

PubMed1999

F-box/WD-repeat proteins pop1p and Sud1p/Pop2p form complexes that bind and direct the proteolysis of cdc18p.

Ubiquitin-dependent proteolysis plays an important role in cell-cycle control [1] [2]. In budding yeast, the protein Skp1p, the cullin-family member Cdc53p, and the F-box/WD-repeat protein Cdc4p form the SCFCdc4p ubiquitin ligase complex, which targets the cyclin-dependent kinase (Cdk) inhibitor Sic1p for proteolysis [3] [4] [5] [6] [7] [8]. Sic1p is recruited to the SCFCdc4p complex by binding to the WD-repeat region of Cdc4p [5] [6], while Skp1p binds to the F-box of Cdc4p [9]. In fission yeast, two distinct Cdc4p-related proteins, Pop1p/Ste16p [10] [11] and the recently identified Sud1p/Pop2p [12], regulate the stability of the replication initiator Cdc18p and the Cdk inhibitor Rum1p. We show here that, despite their structural and functional similarities, the pop1 and pop2 genes fail to complement each other's deletion phenotypes, indicating that they perform non-redundant, but potentially interdependent, functions in proteolysis. Consistent with this hypothesis, Pop1p and Pop2p formed heterooligomeric complexes when overexpressed, and binding of Cdc18p to Pop2p was dependent on Pop1p. The Pop1p-Pop2p interaction was mediated by the amino-terminal domain of Pop2p which, when fused to full-length Pop1p, rescued the phenotype of a Deltapop1Deltapop2 double mutant. Thus, close physical proximity of two distinct F-box/WD-repeat proteins directs proteolysis mediated by the SCFPop ubiquitin ligase complex.

Binding Sites↗

PubMed1995

The G-box: a ubiquitous regulatory DNA element in plants bound by the GBF family of bZIP proteins.

The G-box (CACGTG) is a ubiquitous, cis-acting DNA regulatory element found in plant genomes. Proteins known as G-box factors (GBFs) bind to G-boxes in a context-specific manner, mediating a wide variety of gene expression patterns. We suggest that, as for many biological systems, different combinations of these common elements can lead to diversity and specificity in the regulation of plant gene expression.

Amino Acid Sequence↗