Rok1p is a putative RNA helicase required for rRNA processing.
The synthesis of ribosomes involves many small nucleolar ribonucleoprotein particles (snoRNPs) as transacting factors. Yeast strains lacking the snoRNA, snR10, are viable but are impaired in growth and delayed in the early pre-rRNA cleavages at sites A0, A1, and A2, which lead to the synthesis of 18S rRNA. The same cleavages are inhibited by genetic depletion of the essential snoRNP protein Gar1p. Screens for mutations showing synthetic lethality with deletion of the SNR10 gene or with a temperature-sensitive gar1 allele both identified the ROK1 gene, encoding a putative, ATP-dependent RNA helicase of the DEAD-box family. The ROK1 gene is essential for viability, and depletion of Rok1p inhibits pre-rRNA processing at sites A0, A1, and A2, thereby blocking 18S rRNA synthesis. Indirect immunofluorescence by using a ProtA-Rok1p construct shows the protein to be predominantly nucleolar. These results suggest that Rok1p is required for the function of the snoRNP complex carrying out the early pre-rRNA cleavage reactions.
Venema
J
J
European Molecular Biology Laboratory, Heidelberg, Germany. venema@chem.vu.nl
Bousquet-Antonelli
C
C
Gelugne
J P
JP
Caizergues-Ferrer
M
M
Tollervey
D
D
D000595:Amino Acid Sequence; D002466:Cell Nucleolus / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005656:Fungal Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009687:Nuclear Proteins / Q000235:genetics; D020365:RNA Helicases; D012316:RNA Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012322:RNA Precursors / Q000378:metabolism; D012337:RNA, Ribosomal, 18S / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism; D020718:Ribonucleoproteins, Small Nucleolar; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D016335:Zinc Fingers
D005656:Fungal Proteins; D009687:Nuclear Proteins; D012322:RNA Precursors; D012337:RNA, Ribosomal, 18S; D012342:RNA, Small Nuclear; D017411:Ribonucleoproteins, Small Nuclear; D020718:Ribonucleoproteins, Small Nucleolar; D029701:Saccharomyces cerevisiae Proteins; C073180:GAR1 protein, S cerevisiae; D012316:RNA Nucleotidyltransferases; D020365:RNA Helicases