Document Detail


Identification of proteins whose synthesis is modulated during the cell cycle of Saccharomyces cerevisiae.
MedLine Citation:
PMID:  14582195     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We examined the synthesis and turnover of individual proteins in the Saccharomyces cerevisiae cell cycle. Proteins were pulse-labeled with radioactive isotope (35S or 14C) in cells at discrete cycle stages and then resolved on two-dimensional gels and analyzed by a semiautomatic procedure for quantitating gel electropherogram-autoradiographs. The cells were obtained by one of three methods: (i) isolation of synchronous subpopulations of growing cells by zonal centrifugation.; (ii) fractionation of pulse-labeled steady-state cultures according to cell age; and (iii) synchronization of cells with the mating pheromone, alpha-factor. In confirmation of previous studies, we found that the histones H4, H2A, and H2B were synthesized almost exclusively in the late G1 and early S phases. In addition, we identified eight proteins whose rates of synthesis were modulated in the cell cycle, and nine proteins (of which five, which may well be related, were unstable, with half-lives of 10 to 15 min) that might be regulated in the cell cycle by periodic synthesis, modification, or degradation. Based on the time of maximal labeling in the cell cycle and on experiments with alpha-factor and hydroxyurea, we assigned the cell cycle proteins to two classes: proteins in class I were labeled principally in early G1 phase and at a late stage of the cycle, whereas those in class II were primarily synthesized at times ranging from late G1 to mid S phase. At least one major control point for the cell cycle proteins occurred between "start" and early S phase. A set of stress-responsive proteins was also identified and analyzed. The rates of synthesis of these proteins were affected by certain perturbations that resulted during selection of synchronous cell populations and by heat shock.
Authors:
A T Lörincz; M J Miller; N H Xuong; E P Geiduschek
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Molecular and cellular biology     Volume:  2     ISSN:  0270-7306     ISO Abbreviation:  Mol. Cell. Biol.     Publication Date:  1982 Dec 
Date Detail:
Created Date:  2003-10-29     Completed Date:  2003-11-21     Revised Date:  2010-09-21    
Medline Journal Info:
Nlm Unique ID:  8109087     Medline TA:  Mol Cell Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1532-49     Citation Subset:  IM    
Affiliation:
Department of Biology, University of California at San Diego, La Jolla, California 92093, USA.
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MeSH Terms
Descriptor/Qualifier:
Autoradiography
Carbon Radioisotopes / metabolism
Cell Cycle / physiology*
DNA Replication
Electrophoresis, Gel, Two-Dimensional
Histones / chemistry,  metabolism
Hydroxyurea
Nucleic Acid Synthesis Inhibitors / metabolism
Saccharomyces cerevisiae / cytology,  metabolism*
Saccharomyces cerevisiae Proteins / chemistry,  metabolism*
Sulfur Radioisotopes / metabolism
Grant Support
ID/Acronym/Agency:
GM18686/GM/NIGMS NIH HHS; GM22940/GM/NIGMS NIH HHS; RR00757/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Carbon Radioisotopes; 0/Histones; 0/Nucleic Acid Synthesis Inhibitors; 0/Saccharomyces cerevisiae Proteins; 0/Sulfur Radioisotopes; 127-07-1/Hydroxyurea
Comments/Corrections

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