The transcriptional regulator Hap1p (Cyp1p) is essential for anaerobic or heme-deficient growth of Saccharomyces cerevisiae: Genetic and molecular characterization of an extragenic suppressor that encodes a WD repeat protein.
We report here that Hap1p (originally named Cyp1p) has an essential function in anaerobic or heme-deficient growth. Analysis of intragenic revertants shows that this function depends on the amino acid preceding the first cysteine residue of the DNA-binding domain of Hap1p. Selection of recessive extragenic suppressors of a hap1-hem1- strain allowed the identification, cloning, and molecular analysis of ASC1 (Cyp1 Absence of growth Supressor). The sequence of ASC1 reveals that its ORF is interrupted by an intron that shelters the U24 snoRNA. Deletion of the intron, inactivation of the ORF, and molecular localization of the mutations show unambiguously that it is the protein and not the snoRNA that is involved in the suppressor phenotype. ASC1, which is constitutively transcribed, encodes an abundant, cytoplasmically localized 35-kD protein that belongs to the WD repeat family, which is found in a large variety of eucaryotic organisms. Polysome profile analysis supports the involvement of this protein in translation. We propose that the absence of functional Asc1p allows the growth of hap1-hem1- cells by reducing the efficiency of translation. Based on sequence comparisons, we discuss the possibility that the protein intervenes in a kinase-dependent signal transduction pathway involved in this last function.
Chantrel
Y
Y
Centre de Génétique Moléculaire du Centre National de la Recherche Scientifique, l'Université Pierre et Marie Curie, Gif-sur-Yvette, France.
Gaisne
M
M
Lions
C
C
Verdière
J
J
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D019757:Carbon-Oxygen Lyases; D003001:Cloning, Molecular; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D005656:Fungal Proteins / Q000737:chemistry* / Q000502:physiology; D019204:GTP-Binding Proteins; D005800:Genes, Fungal / Q000235:genetics; D006418:Heme / Q000172:deficiency / Q000494:pharmacology; D007150:Immunohistochemistry; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000502:physiology*; D016366:Open Reading Frames / Q000235:genetics; D011132:Polyribosomes / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D011494:Protein Kinases / Q000502:physiology; D012342:RNA, Small Nuclear / Q000235:genetics; D012270:Ribosomes / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D015398:Signal Transduction / Q000502:physiology; D013489:Suppression, Genetic / Q000235:genetics
C111825:ASC1 protein, S cerevisiae; D048868:Adaptor Proteins, Signal Transducing; D005656:Fungal Proteins; D009687:Nuclear Proteins; D012342:RNA, Small Nuclear; D029701:Saccharomyces cerevisiae Proteins; D006418:Heme; D011494:Protein Kinases; D019204:GTP-Binding Proteins; D019757:Carbon-Oxygen Lyases; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase