Major determinants of the specificity of interaction between small nuclear ribonucleoproteins U1A and U2B" and their cognate RNAs

Scherly, Daniel; Boelens, Wilbert C.; Dathan, Nina A.; Venrooij, Walther J. van; Mattaj, Iain W.

doi:10.1038/345502a0

Cited by 282 publications

(327 citation statements)

References 31 publications

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“…One of the least conserved regions in the RNP-CS RBD is the region preceeding RNP1, referred to as loop 3. Amino acids in loop 3 are thought to be important for determining RNA-binding specificity, and this has been shown experimentally for two proteins, the U1-A and U2-B" snRNP proteins (38 obtained. Interestingly, immunopurification and two-dimensional gel analysis indicated that monoclonal antibody 8A6 recognizes hnRNP F, hnRNP H and the lower molecular weight hnRNP E proteins, revealing that, in addition to their similar nucleic acidbinding properties, these proteins share a common epitope ( Figure 5).…”

Section: Resultsmentioning

confidence: 99%

The hnRNP F protein: unique primary structure, nucleic acid-binding properties, and subcellular localization

1994

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More than 20 different heterogeneous nuclear ribonucleoproteins (hnRNPs) are associated with premRNAs in the nucleus of mammalian cells and these proteins appear to influence pre-mRNA processing and other aspects of mRNA metabolism and transport. The arrangement of hnRNP proteins on pre-mRNAs is likely to be unique for each RNA and may be determined by the different RNA-binding preferences of each of these proteins. hnRNP F (Mr = 53 kD, pi = 6.1) and hnRNP H (Mr = 56 kD, pl = 6.7-7.1) are abundant components of immunopurified hnRNP complexes and they have distinct nucleic acid binding properties. Unlike other hnRNP proteins which display a varying range of affinities for different ribonucleotidehomopolymers and ssDNA, hnRNP F and hnRNP H bind only to poly(rG) in vitro. hnRNP F and hnRNP H were purified from HeLa cells by poly(rG) affinity chromatography and oligonucleotides derived from peptide sequences were used to isolate a cDNA encoding hnRNP F. The predicted amino acid sequence of hnRNP F revealed a novel protein with three repeated domains related to the RNP consensus sequence RNA-binding domain. Monoclonal antibodies produced against bacterially expressed hnRNP F were specific for both hnRNP F and hnRNP H and recognized related proteins in divergent organisms, including in the yeast Saccharomyces cerevisiae. hnRNP F and hnRNP H are thus highly related immunologically and they share identical peptides. Interestingly, immunofluorescence microscopy revealed that hnRNP F and hnRNP H are concentrated in discrete regions of the nucleoplasm, in contrast to the general nucleoplasmic distribution of previously characterized hnRNP proteins. The unique RNA-binding properties, amino acid sequence and distinct intranuclear localization of hnRNP F and hnRNP H make them novel hnRNP proteins that are likely to be important for the processing of RNAs containing guanosine-rich sequences.

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Section: Resultsmentioning

confidence: 99%

The hnRNP F protein: unique primary structure, nucleic acid-binding properties, and subcellular localization

1994

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“…Mutation analysis of the human U2BЉ gene has previously shown which part of the sequence localizes U2BЉ to the nucleus (Scherly et al, 1990;Bentley and Keene, 1991;Kambach and Mattaj, 1994), but not which part of the sequence is responsible for localization to the coiled body. We have shown that fusion of the first N-terminal RNP-80 motif of the potato U2BЉ protein to GFP is sufficient for localization of the fusion protein to the coiled body.…”

Section: Discussionmentioning

confidence: 99%

“…It has been suggested that U2BЉ may not be required for the splicing reaction itself but may have a role in U2 snRNP biogenesis (Mattaj and De Robertis, 1985;Pan and Prives, 1989). The full-length U2BЉ cDNA has been cloned from potato (Simpson et al, 1991) and has been characterized at the molecular level (Simpson et al, 1991, largely by analogy with the human U2BЉ cDNA (Scherly et al, 1990;Bentley and Keene, 1991;Kambach and Mattaj, 1994). The potato U2BЉ protein contains two RNP-80 motifs separated by a central putative nuclear localization signal (NLS).…”

Section: Introductionmentioning

confidence: 99%

“…The RNP motifs consist of 80 -90 amino acids containing two short, highly conserved sequences (RNP1 and RNP2) and are found in numerous proteins involved in RNA processing events (Birney et al, 1993;Mattaj, 1993). It has been shown that only the N-terminal RNP motif of U2BЉ is required for specific binding to U2 snRNA (Kambach and Mattaj, 1994;Simpson et al, 1995) and that the interaction between U2BЉ and U2 snRNA requires the presence of a second protein, U2AЈ (Scherly et al, 1990;Simpson et al, 1995).In the present work we show directly for the first time, by time-lapse confocal fluorescence microscopy, that coiled bodies can move within the nucleus and can coalesce. These findings suggest a transport function for coiled bodies and a mechanism for the observed decrease in coiled body number during the cell cycle.…”

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confidence: 99%

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The Movement of Coiled Bodies Visualized in Living Plant Cells by the Green Fluorescent Protein

Boudonck

Dolan

Shaw

1999

MBoC

132

119

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Coiled bodies are nuclear organelles that contain components of at least three RNAprocessing pathways: pre-mRNA splicing, histone mRNA 3Ј-maturation, and pre-rRNA processing. Their function remains unknown. However, it has been speculated that coiled bodies may be sites of splicing factor assembly and/or recycling, play a role in histone mRNA 3Ј-processing, or act as nuclear transport or sorting structures. To study the dynamics of coiled bodies in living cells, we have stably expressed a U2BЉ-green fluorescent protein fusion in tobacco BY-2 cells and in Arabidopsis plants. Time-lapse confocal microscopy has shown that coiled bodies are mobile organelles in plant cells. We have observed movements of coiled bodies in the nucleolus, in the nucleoplasm, and from the periphery of the nucleus into the nucleolus, which suggests a transport function for coiled bodies. Furthermore, we have observed coalescence of coiled bodies, which suggests a mechanism for the decrease in coiled body number during the cell cycle. Deletion analysis of the U2BЉ gene construct has shown that the first RNP-80 motif is sufficient for localization to the coiled body. INTRODUCTIONThe coiled body was first described by Ramon y Cajal (1903), who called it the "nucleolar accessory body" because of its association with the nucleolus. This nuclear organelle was later reidentified by electron microscopy and renamed the "coiled body" because of its appearance as loosely packed coiled fibrils (Monneron and Bernhard, 1969). Subsequent studies detected coiled bodies in animal and plant nuclei, showing that it is a conserved structure (Moreno Diaz de la Espina et al., 1980;Seite et al., 1982;Schultz, 1990).Coiled bodies have been shown to contain splicing small nuclear ribonucleoproteins (snRNPs) and small nuclear RNAs (snRNAs), a subset of nucleolar components -including fibrillarin, Nopp140, NAP57, and U3 small nucleolar ribonucleoprotein (U3 snoRNP) -and the protein p80 coilin, which has been widely used as a marker for coiled bodies (Lamond and Earnshaw, 1998;Matera, 1998). The function of coiled bodies is still under debate, but several hypotheses have been proposed that are not necessarily mutually exclusive. Because coiled bodies do not contain DNA, nascent pre-mRNA, heterogeneous nuclear RNPs (hnRNPs), or the SC-35 splicing factor, which is required for splicing in vitro, it has been argued that they are not directly involved in transcription and pre-mRNA splicing . However, it has been speculated that coiled bodies may be sites of splicing factor assembly or recycling, may play a role in histone mRNA 3Ј processing (Gall et al., 1995;Lamond and Earnshaw, 1998), or may be involved in all these activities. As coiled bodies are frequently observed at the nucleolar periphery and also in the nucleoplasm and within nucleoli (Malatesta et al. 1994;Ochs et al., 1994), they may also act as nuclear transport or sorting structures. It has been shown recently that coiled bodies are also involved in processing or transport of small nucleolar RNA (snoRNA) pr...

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“…In particular, the fact that both differences are at the loop-3 region, which lies at the N-terminal end of RNP-1 P-sheet, is intriguing. Since the loop-3 and RNP-1 sequences of U 1-A and U2-B" have been shown to determine the binding specificity of these proteins (Scherly et al, 1990;Bentley and Keene, 199 l), the optional addition of 5 amino acids to the RBP9 protein in this region and the 13 extra amino acids of efav at the loop-3 region may confer on them different RNA binding specificities or affinities. In addition, the facts that two related fly proteins, elav and rbp9, have a different expression profile-eelav is required for the early embryo neurogenesis whereas rbp9 is expressed only at the late developmental stages-and that a rbp9 cDNA under the efav promoter failed to rescue the embryonic lethal phenotype of elav (K. M. Yao and K. White, personal communication) are consistent with the idea that these two proteins are involved in different functions rather than having synonymous functions in the CNS development.…”

Section: Discussionmentioning

confidence: 99%

The Drosophila gene rbp9 encodes a protein that is a member of a conserved group of putative RNA binding proteins that are nervous system-specific in both flies and humans

Kim

Baker

1993

J. Neurosci.

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The rbp9 gene of Drosophila melanogaster has been molecularly characterized and shown to be expressed solely in the CNS, where it encodes proteins with three RNA recognition motifs (RRMs). Sequencing of genomic and cDNA clones of rbp9 revealed a complex gene with three alternative promoters as well as alternative patterns of splicing. The deduced amino acid sequence of the RBP9 proteins is highly similar to those of three other nervous system-specific genes, human HuC and HuD and Drosophila elav, which also encode proteins with three RRMs. Developmental Northern analysis revealed that rbp9 is expressed from the late third instar larva through adult stages. The RBP9 protein was detected specifically in nuclei of the nervous system after morphogenesis of the adult CNS in the mid-pupal stage. Thus, the RBP9 protein does not appear until substantially later than rbp9 transcripts are detected. The adult nervous system nuclear-limited expression pattern, the presence of RRMs, and the high similarity to a group of nervous system-specific proteins in flies and humans suggest that rbp9 belongs to a nervous system-specific RRM protein gene subfamily that may participate in the processing of RNAs involved in the development of the CNS.

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Major determinants of the specificity of interaction between small nuclear ribonucleoproteins U1A and U2B" and their cognate RNAs

Cited by 282 publications

References 31 publications

The hnRNP F protein: unique primary structure, nucleic acid-binding properties, and subcellular localization

The hnRNP F protein: unique primary structure, nucleic acid-binding properties, and subcellular localization

The Movement of Coiled Bodies Visualized in Living Plant Cells by the Green Fluorescent Protein

The Drosophila gene rbp9 encodes a protein that is a member of a conserved group of putative RNA binding proteins that are nervous system-specific in both flies and humans

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