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M101564200v1
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Papers In Press, published online ahead of print July 30, 2001
J. Biol. Chem, 10.1074/jbc.M101564200
Submitted on February 20, 2001
Revised on July 30, 2001
Accepted on July 27, 2001

Overexpression of eIF4E in S. cerevisiae causes slow growth and decreased alpha-factor response through alterations in Cln3 expression

Charles Anthony, Qin Zong, and Arrigo De Benedetti

Biochemistry and Molecular Biology, LSU Health Sciences Center - Shreveport, Shreveport, LA 71130

Corresponding Author: adeben{at}lsuhsc.edu

The association of G1 cyclins and Cdc28/CDK catalyzes the cell cycle entry (Start) in budding yeast. Activation of Start is presumed to be triggered by a post-transcriptional increase in CLN3 during early G1. Cells arrested by mating pheromone show a loss of CDK activity caused by transcriptional shutoff of cyclins and/or inhibition by Far1. We report that overexpression of eIF4E (CDC33), a rate limiting translation initiation factor, causes an increase in CLN3 mRNA translation, which results in increased expression of CLN2 and in slow growth and decreased -factor response. This phenotype was abrogated in a cln3 or cln2 background. We isolated the transcription factor MBP1 as a multicopy suppressor of the growth and -factor response defects. Furthermore, elevated MBP1, a transcriptional regulator of cyclins, altered the transcriptional start site in CLN3 mRNA, shifting it 45 nucleotides upstream of the normal. This lengthened 5’ UTR likely reduces translation efficiency and down-regulates CLN3 protein synthesis, thereby correcting for the excess translation promoted by elevated CDC33. In addition, the CLN2 mRNA levels returned to normal. We propose that regulation of translation initiation by CDC33 plays a pivotal role in the activation of Start and cell cycle progression in budding yeast.


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