A human RNA helicase-like protein, HRH1, facilitates nuclear export of spliced mRNA by releasing the RNA from the spliceosome.
Because the nuclear export of mRNA occurs only after the splicing reaction is completed, intron-containing pre-mRNA does not normally appear in the cytoplasm. As a mechanism to secure this, intron-containing RNA is retained in the nucleus via formation of the spliceosome. Therefore, the process of releasing spliced mRNA from the spliceosome after completion of splicing is an essential step for triggering the nuclear export of the spliced mRNA. In budding yeast, RNA helicase-like protein Prp22 is implicated in this process. Here we demonstrate the function of HRH1, a human protein homologous to Prp22, in mammalian cells using dominant-negative HRH1++ mutants (dn-HRH1). dn-HRH1 protein stalls on the spliceosome and prevents release of the spliced RNA from the spliceosome in vitro. Expression of dn-HRH1 in mammalian cells leads to inhibition of splicing and to extensive nuclear export of unspliced pre-mRNA, probably because of the incapability of recycling spliceosome components that normally retain the pre-mRNA in the nucleus. The arginine/serine-rich domain (RS domain) of HRH1, which is missing in Prp22, confers a nuclear localization signal, and appears to facilitate the interaction of HRH1 with the spliceosome. This is the first report on a bona fide mammalian homolog of yeast Prp splicing factor, and also on a mammalian RNA helicase-like splicing factor.
Ohno
M
M
Department of Biophysics, Faculty of Science, Kyoto University, Japan.
Shimura
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002460:Cell Line; D002522:Chlorocebus aethiops; D017931:DNA Primers / Q000737:chemistry; D005799:Genes, Dominant; D006801:Humans; D008969:Molecular Sequence Data; D020365:RNA Helicases; D012316:RNA Nucleotidyltransferases / Q000502:physiology*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D017384:Sequence Deletion; D017381:Spliceosomes / Q000378:metabolism*; D013329:Structure-Activity Relationship
D017931:DNA Primers; D012333:RNA, Messenger; D012316:RNA Nucleotidyltransferases; D020365:RNA Helicases