Dbp5p/Rat8p is a yeast nuclear pore-associated DEAD-box protein essential for RNA export.
To identify Saccharomyces cerevisiae genes important for nucleocytoplasmic export of messenger RNA, we screened mutant strains to identify those in which poly(A)+ RNA accumulated in nuclei under nonpermissive conditions. We describe the identification of DBP5 as the gene defective in the strain carrying the rat8-1 allele (RAT = ribonucleic acid trafficking). Dbp5p/Rat8p, a previously uncharacterized member of the DEAD-box family of proteins, is closely related to eukaryotic initiation factor 4A(eIF4A) an RNA helicase essential for protein synthesis initiation. Analysis of protein databases suggests most eukaryotic genomes encode a DEAD-box protein that is probably a homolog of yeast Dbp5p/Rat8p. Temperature-sensitive alleles of DBP5/RAT8 were prepared. In rat8 mutant strains, cells displayed rapid, synchronous accumulation of poly(A)+ RNA in nuclei when shifted to the non-permissive temperature. Dbp5p/Rat8p is located within the cytoplasm and concentrated in the perinuclear region. Analysis of the distribution of Dbp5p/Rat8p in yeast strains where nuclear pore complexes are tightly clustered indicated that a fraction of this protein associates with nuclear pore complexes (NPCs). The strong mutant phenotype, association of the protein with NPCs and genetic interaction with factors involved in RNA export provide strong evidence that Dbp5p/Rat8p plays a direct role in RNA export.
Snay-Hodge
C A
CA
Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03755, USA.
Colot
H V
HV
Goldstein
A L
AL
Cole
C N
CN
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000201:enzymology; D017931:DNA Primers; D016208:Databases, Factual; D039601:Eukaryotic Initiation Factor-4A; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005800:Genes, Fungal; D005838:Genotype; D051379:Mice; D008969:Molecular Sequence Data; D009685:Nuclear Envelope / Q000201:enzymology; D010448:Peptide Initiation Factors / Q000737:chemistry; D019151:Peptide Library; D020365:RNA Helicases; D012316:RNA Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*; D012568:Schizosaccharomyces / Q000201:enzymology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D005656:Fungal Proteins; D010448:Peptide Initiation Factors; D019151:Peptide Library; D012333:RNA, Messenger; D011994:Recombinant Proteins; D039601:Eukaryotic Initiation Factor-4A; D012316:RNA Nucleotidyltransferases; D020365:RNA Helicases