CLN3 expression is sufficient to restore G1-to-S-phase progression in Saccharomyces cerevisiae mutants defective in translation initiation factor eIF4E.
The essential cap-binding protein (eIF4E) of Saccharomyces cerevisiae is encoded by the CDC33 (wild-type) gene, originally isolated as a mutant, cdc33-1, which arrests growth in the G1 phase of the cell cycle at 37 degrees C. We show that other cdc33 mutants also arrest in G1. One of the first events required for G1-to-S-phase progression is the increased expression of cyclin 3. Constructs carrying the 5'-untranslated region of CLN3 fused to lacZ exhibit weak reporter activity, which is significantly decreased in a cdc33-1 mutant, implying that CLN3 mRNA is an inefficiently translated mRNA that is sensitive to perturbations in the translation machinery. A cdc33-1 strain expressing either stable Cln3p (Cln3-1p) or a hybrid UBI4 5'-CLN3 mRNA, whose translation displays decreased dependence on eIF4E, arrested randomly in the cell cycle. In these cells CLN2 mRNA levels remained high, indicating that Cln3p activity is maintained. Induction of a hybrid UBI4 5'-CLN3 message in a cdc33-1 mutant previously arrested in G1 also caused entry into a new cell cycle. We conclude that eIF4E activity in the G1-phase is critical in allowing sufficient Cln3p activity to enable yeast cells to enter a new cell cycle.
Danaie
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P
Institute for Biochemistry and Molecular Biology, University of Berne, Bühlstrasse 28, CH-3012 Bern, Switzerland.
Altmann
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M
Hall
M N
MN
Trachsel
H
H
Helliwell
S B
SB
D020121:5' Untranslated Regions / Q000235:genetics; D016213:Cyclins / Q000235:genetics / Q000502:physiology*; D039561:Eukaryotic Initiation Factor-4E; D005434:Flow Cytometry; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology*; D016193:G1 Phase; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal / Q000235:genetics / Q000502:physiology; D017930:Genes, Reporter; D009154:Mutation; D010448:Peptide Initiation Factors / Q000235:genetics / Q000502:physiology*; D014176:Protein Biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins
D020121:5' Untranslated Regions; C082721:CLN2 protein, S cerevisiae; C079895:CLN3 protein, S cerevisiae; D016213:Cyclins; D039561:Eukaryotic Initiation Factor-4E; D005656:Fungal Proteins; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins