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PubMed2002

Cooperative E-box regulation of human GLI1 by TWIST and USF.

Sonic hedgehog signaling plays a critical role in vertebrate patterning, and signaling defects are associated with severe birth defects and cancer in man. GLI1 encodes a critical transcription activator in this pathway. GLI1 is expressed in human basal cell carcinomas and sarcomas. Despite the significance of the GLI1 gene in human disease, few immediate upstream regulators of GLI1 expression are known. We previously demonstrated that a 5' region, including 5' flanking sequence, an untranslated exon, and 425 bp of the first intron, regulates the human GLI1 gene. Here we show that inactivating mutations in E-box, GC box, AP-2, GATA, GSG, PuF, and Zeste sites identified three critical regulatory elements, including a GC box that binds Sp1 and two intronic E-boxes that bind USF proteins or Twist. Expression of Twist but not a frame shift mutation of Twist activates the wild-type human GLI1 regulatory sequences but not with inactivating mutations of the E-boxes. Twist activates GLI1 reporter expression through E-box +482 but requires binding of USF proteins to E-box +157. Twist mutations cause human birth defects and Twist is overexpressed in many rhabdomyosarcomas, suggesting that one of Twist's primary roles is the regulation of GLI1.

Cloning, Molecular↗

PubMed1986

Stage-specific expression of a homeo box-containing gene in the non-segmented sea urchin embryo.

Hybridization of Drosophila homeo box DNA probes to Southern transfers of genomic DNA from the Hawaiian sea urchin Tripneustes gratilla has revealed that the sea urchin genome contains at least five homeo boxes. Examination of the DNA from several individuals shows that the sequences flanking these homeo boxes exhibit little restriction fragment length polymorphism, indicating they are more highly conserved than the majority of sea urchin DNA. Several clones in a T. gratilla genomic DNA library which hybridized with Drosophila homeo box probes were identified, and one found to be transcribed during embryogenesis was selected for further study. Southern transfer hybridizations showed the cloned gene to be single-copy. DNA sequencing of the sea urchin gene defined a homeo box 70-73% homologous to the Drosophila homeo box probes and an encoded homeo domain 78-88% homologous to those encoded by the probes. Hybridization of DNA probes from the sea urchin homeo box-containing gene to Northern transfers of embryonic RNA demonstrated that the gene produces two transcripts of 6.9 kb and 7.7 kb. Transcripts first accumulate at blastula stage and increase to a maximum level at gastrula stage before decreasing considerably in abundance by pluteus stage.

Amino Acid Sequence↗

PubMed1988

Regulation of initiation of the chromosomal replication by DnaA-boxes in the origin region of the Bacillus subtilis chromosome.

A gene homologous to the Escherichia coli dnaA gene and two flanking 'regulatory' regions which contain nine and four DnaA-boxes respectively, are located in the replication origin region of the Bacillus subtilis chromosome. Attempts to isolate an autonomously replicating fragment from these 'regulatory' regions in order to identify oriC have been unsuccessful because the DnaA-box-containing regions strongly inhibited plasmid transformation particularly when inserted into a high-copy number plasmid pUB110. Using two plasmids differing in copy number, the two regions were subdivided into three regions, A, B and C, each containing five, four and four DnaA-boxes respectively, which differed in level of inhibition of transformation. Region C is downstream of the 'dnaA' gene and inhibits transformation in high-copy but not in low-copy number plasmids. When a part of the DnaA-boxes was deleted from the incompatible plasmids, they became transformable and produced slow-growing transformants in which the initiation frequency of chromosomal replication was selectively reduced. Fast-growing revertants were found containing the same number of plasmids as the parent but with single base changes in the DnaA-boxes. These mutations were in the most highly conserved bases of the DnaA-box sequence. This indicates that a sequence-specific interaction of the DnaA-box, probably with the B. subtilis DnaA protein is responsible for the observed incompatibility and thus appears to be involved in control of initiation frequency of the chromosomal replication.

Bacillus subtilis↗

PubMed1994

The DNA sequence specificity of HMG boxes lies in the minor wing of the structure.

To establish the basis of sequence-specific DNA recognition by HMG boxes we separately transferred the minor and major wings from the sequence-specific HMG box of TCF1 alpha into their equivalent position in the non-sequence-specific box 2 of HMG1. Thus chimera THT1 contains the minor wing (of 11 N-terminal and 25 C-terminal residues) from the HMG box of TCF1 alpha and the major wing (the 45 residue central section) from HMG1 box 2, whilst the situation is reversed in chimera HTH1. The structural integrity of the two chimeric proteins was established by CD, NMR and their binding to four-way junction DNA. Gel retardation and circular permutation assays showed that only chimera THT1, containing the TCF1 alpha minor wing, formed a sequence-specific complex and bent the DNA. The bend angle was estimated to be 59 degrees for chimera THT1 and 52 degrees for the HMG box of TCF1 alpha. Our results, in combination with mutagenesis and other data, suggests a model for the DNA binding of HMG boxes in which the N-terminal residues and part of helix 1 contact the minor groove on the outside of a bent DNA duplex.

Amino Acid Sequence↗

PubMed2005

Upstream stimulatory factor regulates E box-dependent PAI-1 transcription in human epidermal keratinocytes.

Certain growth factors (e.g., TGF-beta1) initiate a "plastic" response in human keratinocytes (HaCaT cells) characterized by changes in gene expression and increased cell motility. While microarray analyses identified a number of involved genes, plasminogen activator inhibitor type 1 (PAI-1) is among the subset most highly responsive to TGF-beta1. Previous antisense attenuation of PAI-1 synthesis confirmed an essential role for this protease inhibitor in cell motility (Providence et al., 2002, J Cell Sci 115:3767-3777; Providence and Higgins, 2004, J Cell Physiol 200:297-308). It was important, therefore, to clarify molecular mechanisms underlying PAI-1 expression control in human keratinocytes. A consensus E box motif (5'-CACGTG-3') at nucleotides -566 to -561 in the PE2 region of the PAI-1 gene was required for TGF-beta1-induced transcription of a PAI-1 promoter-driven luceriferase reporter. Truncation of the PE2 E box or mutation of the CACGTG hexanucleotide to CAATTG inhibited growth factor-stimulated promoter function confirming the importance of this site in inducible expression. A similar mutation at the PE1 E box (nucleotides -682 to -677), in contrast, did not result in reduced luciferase activity. Competing CACGTG-containing DNAs, regardless of the presence or absence of PAI-1-specific flanking sequences or lacking accessory sequences (i.e., Smad-binding sites, AAT trinucleotide spacer), inhibited complex formation between HaCaT cell nuclear factors and a 45-mer PE2 region probe. A deoxyoligonucleotide that differed from the consensus E box by a CG --> AT substitution (the same base change incorporated into the PAI-1p806-lucerifase reporter by site-directed mutagenesis) but with random (i.e., non-PAI-1) flanking sequences also failed to compete with the PE2 region probe for protein binding whereas the same construct with an intact CACGTG motif was an effective competitor. The major protein/DNA interactions in the PE2 segment, therefore, are E box-dependent. USF-1, a member of the upstream stimulatory factor family, bound the PE2 construct suggesting a role for USF proteins in E box residence and PAI-1 gene expression. Chromatin immunoprecipitation, using primers designed to amplify a 300-bp PE2-associated promoter fragment and containing no other E box motifs except the target CACGTG at nucleotides -566 to -561, confirmed that this site was occupied by USF-1 or a USF-1-containing complex in both quiescent and TGF-beta1-stimulated cells. Transfection of a dominant-negative USF construct effectively attenuated serum- and TGF-beta1-induced PAI-1 synthesis as well as TGF-beta1-stimulated Matrigel barrier invasion. Dominant-negative USF-expressing keratinocytes, moreover, specifically had a reduced capacity for Matrigel barrier invasion. USF elements, therefore, are important regulators of growth factor-initiated PAI-1 transcription (as predicted from the identification of PAI-1 as a direct USF target gene) and the associated epithelial migratory response.

Base Sequence↗

PubMed2001

DnaA boxes in the P1 plasmid origin: the effect of their position on the directionality of replication and plasmid copy number.

The DnaA protein is essential for initiation of DNA replication in a wide variety of bacterial and plasmid replicons. The replication origin in these replicons invariably contains specific binding sites for the protein, called DnaA boxes. Plasmid P1 contains a set of DnaA boxes at each end of its origin but can function with either one of the sets. Here we report that the location of origin-opening, initiation site of replication forks and directionality of replication do not change whether the boxes are present at both or at one of the ends of the origin. Replication was bidirectional in all cases. These results imply that DnaA functions similarly from the two ends of the origin. However, origins with DnaA boxes proximal to the origin-opening location opened more efficiently and maintained plasmids at higher copy numbers. Origins with the distal set were inactive unless the adjacent P1 DNA sequences beyond the boxes were included. At either end, phasing of the boxes with respect to the remainder of the origin influenced the copy number. Thus, although the boxes can be at either end, their precise context is critical for efficient origin function.

Bacterial Proteins↗

PubMed1993

Is the use of boxed gloves in an intensive care unit safe?

PURPOSE: To identify the type, rate, burden, and pattern of contamination of boxed, clean but nonsterile gloves in our intensive care unit (ICU). MATERIALS AND METHODS: The fingertips of the first, middle, and last two pairs of gloves in 29 boxes in routine service in our ICU were cultured. The first of each of these three sets were removed aseptically, the second in a routine fashion. RESULTS: We found 16 of 29 (55%) first pairs removed aseptically to be contaminated with a mean bioburden of 1.8 colony-forming units (CFU). The percentage contamination and bioburden did not change significantly with position in the box. Use of routine compared with strict aseptic technique increased the rate of contamination by only 11% (95% confidence interval [CI] -0.05 to +0.27 percentage points) and bioburden by only a mean of 3.4 colonies per pair (CI -0.51 to +4.90 CFU). The length of the time the boxes were open and in use was unrelated to whether the final aseptically removed pair was sterile or contaminated. The predominant organisms were coagulase-negative staphylococci. CONCLUSIONS: One half the pairs of latex examination gloves in our ICU were sterile despite repeated barehanded access to the boxes. Those contaminated exhibited a small bioburden of low pathogenic potential. No pattern of contamination or unsafe duration of box use were observed. The use of boxed, clean, nonsterile gloves appears safe for routine use in an ICU.

Bacterial Infections↗

PubMed1995

Evolutionary conservation in the DNA-binding and -bending properties of HMG-boxes from SRY proteins of primates.

Mammalian sex determination is caused by the Y-chromosome gene SRY, which encodes a protein containing a DNA-binding domain (HMG-box) of about 70 amino acids (aa). The HMG-box is very conserved in a wide variety of mammals; conversely, the flanking non-box regions show a high degree of aa sequence divergence, even between closely related species. The HMG-box of human SRY binds sequence-specifically to linear DNA and produces a sharp bend; it also interacts with high affinity to kinked DNA structures irrespective of their sequences. Point mutations associated with sex reversal in XY human females fall within the HMG-box and either affect the affinity for DNA or modify the geometry of the DNA-protein complex. Here, we show that the DNA-binding and -bending properties of the HMG-boxes of SRY from human and seven different primates are extremely similar to each other. Together with other data, this suggests that the inability of mouse and human SRY to substitute for each other is due to differences in the conserved HMG-box, rather than the non-conserved flanking sequences.

Amino Acid Sequence↗

PubMed2006

Two GC-rich boxes in huC promoter play distinct roles in controlling its neuronal specific expression in zebrafish embryos.

HuC, a vertebrate ortholog of Drosophila elav gene, encodes an RNA binding protein and is involved in early neurogenesis. Zebrafish huC is expressed in distinct neurons, including Rohon-Beard (RB) sensory neurons, interneurons and motoneurons, during primary neurogenesis, and in all neurons later during secondary neurogenesis. In this study, we identify two GC-rich box elements, proximal GC (p-GC) box from -172 to -149 and distal GC (d-GC) box from -218 to -208, in zebrafish huC promoter. Using transgenic approach, we demonstrate that deletion of the p-GC box from the promoter results in loss of expression of the reporter GFP in neurons while deletion of the d-GC box leads to GFP expression only in dorsal RB sensory neurons. These results suggest that the p-GC box alone confers transcriptional activity of huC promoter in primary RB neurons and the d-GC is required for huC transcription in the full spectrum of spinal cord neurons. Further studies are needed to identify specific Sp1-like transcription factors that bind to these GC boxes and activate huC transcription.

Animals↗

PubMed2006

The "black box" warning and allergy drugs.

In the past years, several drugs commonly used by allergy specialists have received a "black box" warning added to their package insert at the direction of the Food and Drug Administration (FDA). A "black box" warning is the highest level of 5 possible warning categories found in the package insert. The FDA has never articulated the basis for "black box" warnings. They generally appear to be based on clinical data, but occasionally can be based on serious animal toxicity. In the last several years, several drugs commonly used by allergists have received recommendations for "black box" warnings. Long-acting beta-agonists (salmeterol and formoterol) received "black box" warnings because of reports of the occurrences of severe asthma exacerbations in some patients with asthma, with some associated death. Topical calcineurin inhibitors (tacrolimus and pimecrolimus) received a recommendation for application of a "black box" warnings because of a possible increase of cancer developing in patients taking these drugs. Although the addition of a "black box" warning was recommended by the FDA Pediatric Advisory Committee for these 2 topical agents, the FDA has not yet implemented this warning. Informed consent principles require that a patient be adequately informed of the risks (among other components) of any recommended treatment. The risks, as described, of the long-acting beta-agonists and topical immunosuppressants should be presented to the patients to aid them in deciding whether they are willing to take these drugs when recommended by their physician.

Adrenergic beta-Agonists↗

PubMed2000

Preference for various nest box designs in farmed silver foxes (Vulpes vulpes) and blue foxes (Alopex lagopus).

Nest box choice experiments were carried out outside the breeding season on adult silver and blue fox vixens with no previous permanent nest box experience. Nest boxes were varied in height of placement, number of rooms, presence of entrance room or platform and light conditions. Only one parameter was varied in any one experiment. Both fox species clearly preferred an elevated multi-room nest box; while silver foxes showed preference for boxes supplied with a platform, blue foxes preferred boxes with an entrance room. There was no significant box preference with respect to light conditions. The possible welfare implications of the preferences are discussed.

Journal Article↗

PubMed2000

Isolation and characterisation of the carnation floral-specific MADS box gene, CMB2.

The cDNA clone KD81, was isolated from a carnation petal cDNA library based on its strong differential expression in petals compared with leaves. The deduced amino acid sequence of KD81 indicated high homology with members of the MADS box family of transcription factors. Identified within the deduced amino acid sequence are two conserved domains; an N-terminal, MADS box and a central, K box. The gene encoding KD81 was termed Carnation MADS Box gene 2 (CMB2). Expression of CMB2 is floral-specific and in petal, transcripts were persistent from the initial stages of development through flower opening. Transcripts were not detected in vegetative tissues. The CMB2 protein is most homologous to TDR6 from tomato, the product of the petal and stamen identity gene DEFICIENS (DEFA), and several DEFA homologues including SLM3, STDEF, PMADS1 and APETALA3. Southern blot analysis indicated that CMB2 is present as a single copy within the carnation genome. Characterisation of a genomic clone encoding CMB2, revealed the molecular structure of CMB2 to be consistent with that reported for other plant MADS box genes. Analysis of the CMB2 promoter sequence revealed the presence of two putative cis-acting elements known as serum response elements (SREs). These elements are proposed as the target for MADS box domain binding and may be involved in the regulation/autoregulation of gene expression. CMB2 represents the first reported isolation of a MADS box gene from carnation.

Journal Article↗

PubMed1997

Box tilt and knee motions in manual lifting: two differential factors in expert and novice workers.

OBJECTIVE: It was the objective of this study to investigate the kinematic and kinetic effects of two specific handling factors that differentiate expert and novice workers, namely the level of knee flexion and box tilt. DESIGN: Seven inexperienced subjects were required to lift a 12-kg box in the sagittal plane using three different strategies: (1) reduced knee flexion and a backward box tilt (more typical of experts); (2) large knee flexion and a backward box tilt; (3) large knee flexion and no box tilt (more typical of novices). BACKGROUND: The lifting techniques of highly skilled workers differ substantially from those of novices but only limited information is available to compare their biomechanical differences. METHODS: The methods included dynamic segmental analyses to calculate the net moments at all body joints and a planar single-muscle equivalent to estimate compression loadings at L5/S1; total work and joint work distribution were calculated using the integration of joint power. The 3-D kinematic data were acquired with three video cameras and force data were obtained with one AMTI force platform which were synchronized with the film data. RESULTS: Box tilt reduced the load trajectory and loadings on the lower back and shoulders; a reduced knee flexion affected body posture and reduced mechanical work and loadings on all body joints. The combination of these two factors, box tilt and reduced knee flexion, showed added effects for almost all variables. CONCLUSIONS: These two factors proper to experts were, in this context, biomechanically more advantageous.

Journal Article↗

PubMed2000

The check box: determining pregnancy status to improve maternal mortality surveillance.

OBJECTIVE: More than half of pregnancy-related deaths are not identified through routine surveillance methods. The purpose of this study was to evaluate the effectiveness of the pregnancy check box on death certificates in ascertaining pregnancy-related deaths. METHODS: Data derived from the Centers for Disease Control and Prevention's ongoing Pregnancy Mortality Surveillance System were used to identify states that included a check box on the death certificate in 1991 and 1992. Death certificates from those states were evaluated to determine the number and proportion of pregnancy-related deaths identified by a marked check box. Characteristics of death were also examined. RESULTS: Sixteen states and New York City included a check box or question specifically asking about pregnancy of the decedent. Of the 425 pregnancy-related deaths identified in the 17 reporting areas, 124 (29%) were determined to be pregnancy-related deaths only because of the pregnancy status information provided in the check box. The proportion of deaths identified only by a marked check box ranged from less than 5% for four states to 40% or more for seven states. CONCLUSIONS: The availability of pregnancy status information on death certificates is a simple and effective aid in ascertaining a pregnancy-related death, when no other indicators of pregnancy appear on the death certificate. Routine use of the pregnancy check box for all states would lead to substantially increased classification of maternal deaths and more accurate classification of the causes of and risk factors for maternal deaths.

Death Certificates↗

PubMed1998

The helC gene encodes a putative DEAD-box RNA helicase required for development in Dictyostelium discoideum.

DEAD-box RNA helicases, defined by the sequence Asp-Glu-Ala-Asp (DEAD, in single-letter amino-acid code), regulate RNA unwinding and secondary structure in an ATP-dependent manner in vitro [1] and control mRNA stability and protein translation. Both yeast and mammals have large families of DEAD-box proteins, many of unknown function. We have disrupted a Dictyostelium discoideum gene, helC, which encodes helicase C, a member of the DEAD-box family of RNA helicases that shows strong homology to the product of the essential Saccharomyces cerevisiae gene dbp5 [2] and to related helicases in mouse and Schizosaccharomyces pombe. The HelC protein also shows weaker homology to the translation initiation factor elF-4a. Other DEAD-box-containing proteins, which are less closely related to HelC, have been implicated in developmental roles in Drosophila [3] and Xenopus laevis; one example is the Xenopus Vasa-like protein (XVLP) [4-6]. In Drosophila and Xenopus, Vasa and XVLP, respectively, are required for the establishment of tissue polarity during development. In yeast, DEAD-box helicases such as Prp8 [7] are components of the spliceosome and connect pre-mRNA splicing with the cell cycle. Disruption of the helC gene in D. discoideum led to developmental asynchrony, failure to differentiate and aberrant morphogenesis. We postulate that one reason for the existence of large families of homologous DEAD-box proteins in yeast, mammals and Dictyostelium could be that some DEAD-box proteins have developmentally specific roles regulating protein translation or mRNA stability.

Animals↗

PubMed2003

The major clades of MADS-box genes and their role in the development and evolution of flowering plants.

MADS-box genes encode a family of transcription factors which control diverse developmental processes in flowering plants ranging from root to flower and fruit development. Sequencing of (almost) the complete Arabidopsis genome enabled the identification of (almost) all of the Arabidopsis MADS-box genes. MADS-box genes have been divided in two large groups, termed type I and type II genes. The type II genes comprise the MEF2-like genes of animals and fungi and the MIKC-type genes of plants. The majority of MIKC-type genes are of the MIKC(c)-type, which includes all plant MADS-box genes for which expression patterns or mutant phenotypes are known. By phylogeny reconstruction, almost all of the MIKC(c)-type genes can be subdivided into 12 major gene clades, each clade comprising 1-6 paralogs from Arabidopsis and putative orthologs from other seed plants. Here we first briefly describe the deep branching of the MADS-box gene tree to place the MIKC(c)-type genes into an evolutionary context. For every clade of MIKC(c)-type genes we then review what is known about its members from Arabidopsis and well-studied members from other phylogenetically informative plant species. By gene sampling and phylogeny reconstructions we provide minimal estimates for the ages of the different clades. It turns out that 7 of the 12 major gene clades, i.e., AG-, AGL6-, AGL12-, DEF+GLO- (B), GGM13- (B(s)), STMADS11- and TM3-like genes very likely existed already in the most recent common ancestor of angiosperms and gymnosperms about 300MYA. Three of the other clades, i.e., AGL2-, AGL17-, and SQUA-like genes, existed at least already in the most recent common ancestor of monocots and eudicots about 200 MYA. Only for two gene clades, AGL15-like genes (2 genes in Arabidopsis) and FLC-like genes (6 genes) members from plants other than Brassicaceae have not been reported yet. Similarly, only one ancient clade known from other flowering plant species, TM8-like genes, is not represented in Arabidopsis. These findings reveal that the diversity of MADS-box genes in Arabidopsis is rather ancient and representative for other flowering plants. Our studies may thus help to predict the set of MADS-box genes in all other flowering plants, except for relatively young paralogs. For the different gene clades we try to identify ancestral and derived gene functions and review the importance of these clades for seed plant development and evolution. We put special emphasis on gene clades for which insights into their importance has rapidly increased just recently.

Arabidopsis↗

PubMed2000

Box C/D snoRNA-associated proteins: two pairs of evolutionarily ancient proteins and possible links to replication and transcription.

The eukaryotic nucleolus contains a diverse population of small nucleolar RNAs (snoRNAs) essential for ribosome biogenesis. The box C/D snoRNA family possesses conserved nucleotide boxes C and D that are multifunctional elements required for snoRNA processing, snoRNA transport to the nucleolus, and 2'-O-methylation of ribosomal RNA. We have previously demonstrated that the assembly of an snoRNP complex is essential for processing the intronic box C/D snoRNAs and that specific nuclear proteins associate with the box C/D core motif in vitro. Using a box C/D motif derived from mouse U14 snoRNA, we have now affinity purified and defined four mouse proteins that associate with this minimal RNA substrate. These four proteins consist of two protein pairs: members of each pair are highly related in sequence. One protein pair corresponds to the essential yeast nucleolar proteins Nop56p and Nop58p. Affinity purification of mouse Nop58 confirms observations made in yeast that Nop58 is a core protein of the box C/D snoRNP complex. Isolation of Nop56 using this RNA motif defines an additional snoRNP core protein. The second pair of mouse proteins, designated p50 and p55, are also highly conserved among eukaryotes. Antibody probing of nuclear fractions revealed a predominance of p55 and p50 in the nucleoplasm, suggesting a possible role for the p50/p55 pair in snoRNA production and/or nucleolar transport. The reported interaction of p55 with TATA-binding protein (TBP) and replication A protein as well as the DNA helicase activity of p55 and p50 may suggest the coordination of snoRNA processing and snoRNP assembly with replication and/or transcriptional events in the nucleus. Homologs for both snoRNA-associated protein pairs occur in Archaea, strengthening the hypothesis that the box C/D RNA elements and their interacting proteins are of ancient evolutionary origin.

Amino Acid Motifs↗

PubMed2006

Thermodynamics of the fragile X mental retardation protein RGG box interactions with G quartet forming RNA.

Fragile X syndrome, the most common form of inherited mental retardation, is the result of an unstable expansion of a CGG trinucleotide repeat in the 5' UTR of the fragile X mental retardation-1 (FMR1) gene. The abnormal hypermethylation of the expanded CGG repeats causes the transcriptional silencing of the FMR1 gene and, consequently, the loss of the fragile X mental retardation protein (FMRP). FMRP is an RNA binding protein that binds to G quartet forming RNA using its RGG box motif. In this study we have performed a thermodynamic analysis of the interactions between the FMRP RGG box domain and Sc1, an RNA molecule which had been previously shown to be bound with high affinity by both the full-length FMRP and by its RGG box domain. We have determined that the association between the FMRP RGG box and Sc1 RNA is dominated by hydrophobic and hydrogen bond interactions, with minor contributions from electrostatic interactions, and that the FMRP RGG box binding increases the stability of the G quartet RNA structure significantly. Interestingly, we found that the G quartet recognition is necessary but not sufficient for the FMRP RGG box binding to this RNA target, indicating that additional interactions of the peptide, possibly with the stem and/or stem-G quartet junction region, are required. Our results also indicate that the G quartet RNA recognition is not a general feature of the RGG box motif but rather carries some sequence, protein and/or RNA, specificity.

Amino Acid Sequence↗