Identification of a putative RNA helicase (HRH1), a human homolog of yeast Prp22.
In the budding yeast Saccharomyces cerevisiae, a number of PRP genes known to be involved in pre-mRNA processing have been genetically identified and cloned. Three PRP genes (PRP2, PRP16, and PRP22) were shown to encode putative RNA helicases of the family of proteins with DEAH boxes. However, any such splicing factor containing the helicase motifs in vertebrates has not been identified. To identify human homologs of this family, we designed PCR primers corresponding to the highly conserved region of the DEAH box protein family and successfully amplified five cDNA fragments, using HeLa poly(A)+ RNA as a substrate. One fragment, designated HRH1 (human RNA helicase 1), is highly homologous to Prp22, which was previously shown to be involved in the release of spliced mRNAs from the spliceosomes. Expression of HRH1 in a S. cerevisiae prp22 mutant can partially rescue its temperature-sensitive phenotype. These results strongly suggest that HRH1 is a functional human homolog of the yeast Prp22 protein. Interestingly, HRH1 but not Prp22 contains an arginine- and serine-rich domain (RS domain) which is characteristic of some splicing factors, such as members of the SR protein family. We could show that HRH1 can interact in vitro and in the yeast two-hybrid system with members of the SR protein family through its RS domain. We speculate that HRH1 might be targeted to the spliceosome through this interaction.
Ono
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Y
Department of Biophysics, Faculty of Science, Kyoto University, Japan.
Ohno
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M
Shimura
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D000595:Amino Acid Sequence; D001483:Base Sequence; D053487:DEAD-box RNA Helicases; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D020365:RNA Helicases; D012316:RNA Nucleotidyltransferases / Q000235:genetics*; D012322:RNA Precursors / Q000378:metabolism; D012326:RNA Splicing / Q000235:genetics; D000072260:RNA Splicing Factors; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D017381:Spliceosomes / Q000378:metabolism
D017931:DNA Primers; D004271:DNA, Fungal; D005656:Fungal Proteins; D012322:RNA Precursors; D000072260:RNA Splicing Factors; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; D012316:RNA Nucleotidyltransferases; C067135:PRP22 protein, S cerevisiae; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases