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The budding yeast Saccharomyces cerevisiae CLN1, CLN2, and CLN3 genes encode functionally redundant G1 cyclins required for cell cycle initiation. CLN1 and CLN2 mRNAs accumulate periodically throughout the cell cycle, peaking in late G1. We show that cell cycle-dependent fluctuation in CLN2 mRNA is regulated at the level of transcriptional initiation. Mutational analysis of the CLN2 promoter revealed that the major cell cycle-dependent upstream activating sequence (UAS) resides within a 100-bp fragment. This UAS contains three putative SWI4-dependent cell cycle boxes (SCBs) and two putative MluI cell cycle boxes (MCBs). Mutational inactivation of these elements substantially decreased CLN2 promoter activity but failed to eliminate periodic transcription. Similarly, inactivation of SWI4 decreased CLN2 transcription without affecting its periodicity. We have identified a second UAS in the CLN2 upstream region that can promote cell cycle-dependent transcription with kinetics similar to that of the intact CLN2 promoter. Unlike the major CLN2 UAS, this newly identified UAS promotes transcription in cells arrested in G1 by inactivation of cdc28. This novel UAS is both necessary and sufficient for regulated transcription driven by a CLN2 promoter lacking functional SCBs and MCBs. Although this UAS itself contains no SCBs or MCBs, its activity is dependent upon SWI4 function. The characteristics of this novel UAS suggest that it might have a role in initiating CLN2 expression early in G1 to activate the positive feedback loop that drives maximal Cln accumulation.

Original publication

DOI

10.1128/mcb.14.7.4788

Type

Journal article

Journal

Molecular and Cellular Biology

Publication Date

07/1994

Volume

14

Pages

4788 - 4801

Addresses

Scripps Research Institute, La Jolla, California 92037.

Keywords

Saccharomyces cerevisiae, Cyclins, DNA-Binding Proteins, Fungal Proteins, Saccharomyces cerevisiae Proteins, Transcription Factors, RNA, Messenger, DNA Primers, Restriction Mapping, Mutagenesis, Site-Directed, DNA Mutational Analysis, Cell Cycle, Gene Expression, Transcription, Genetic, Gene Expression Regulation, Fungal, Base Sequence, Regulatory Sequences, Nucleic Acid, Kinetics, Genotype, Genes, Fungal, Plasmids, Time Factors, Molecular Sequence Data, Promoter Regions, Genetic