TY - JOUR
T1 - Relationships between Cell Cycle Regulator Gene Copy Numbers and Protein Expression Levels in Schizosaccharomyces pombe
AU - Chino, Ayako
AU - Makanae, Koji
AU - Moriya, Hisao
N1 - Funding Information:
We would like to thank Masataka Sasabe and Reiko Kintaka (Okayama University) for valuable discussions. We thank the Yeast Genetic Resource Center, Japan (supported by the National BioResource Project) for providing S. pombe strain FY7652.
PY - 2013/9/3
Y1 - 2013/9/3
N2 - We previously determined the copy number limits of overexpression for cell division cycle (cdc) regulatory genes in the fission yeast Schizosaccharomyces pombe using the "genetic tug-of-war" (gTOW) method. In this study, we measured the levels of tandem affinity purification (TAP)-tagged target proteins when their copy numbers are increased in gTOW. Twenty analyzed genes showed roughly linear correlations between increased protein levels and gene copy numbers, which suggested a general lack of compensation for gene dosage in S. pombe. Cdc16 and Sid2 protein levels but not their mRNA levels were much lower than that expected by their copy numbers, which suggested the existence of a post-transcriptional down regulation of these genes. The cyclin Cig1 protein level and its mRNA level were much higher than that expected by its copy numbers, which suggested a positive feedback mechanism for its expression. A higher Cdc10 protein level and its mRNA level, probably due to cloning its gene into a plasmid, indicated that Cdc10 regulation was more robust than that previously predicted.
AB - We previously determined the copy number limits of overexpression for cell division cycle (cdc) regulatory genes in the fission yeast Schizosaccharomyces pombe using the "genetic tug-of-war" (gTOW) method. In this study, we measured the levels of tandem affinity purification (TAP)-tagged target proteins when their copy numbers are increased in gTOW. Twenty analyzed genes showed roughly linear correlations between increased protein levels and gene copy numbers, which suggested a general lack of compensation for gene dosage in S. pombe. Cdc16 and Sid2 protein levels but not their mRNA levels were much lower than that expected by their copy numbers, which suggested the existence of a post-transcriptional down regulation of these genes. The cyclin Cig1 protein level and its mRNA level were much higher than that expected by its copy numbers, which suggested a positive feedback mechanism for its expression. A higher Cdc10 protein level and its mRNA level, probably due to cloning its gene into a plasmid, indicated that Cdc10 regulation was more robust than that previously predicted.
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U2 - 10.1371/journal.pone.0073319
DO - 10.1371/journal.pone.0073319
M3 - Article
C2 - 24019917
AN - SCOPUS:84883373902
SN - 1932-6203
VL - 8
JO - PloS one
JF - PloS one
IS - 9
M1 - e73319
ER -