Regulation of CLB6 expression by the cytoplasmic deadenylase Ccr4 through its coding and 3' UTR regions

PLoS One. 2022 May 6;17(5):e0268283. doi: 10.1371/journal.pone.0268283. eCollection 2022.

Abstract

RNA stability control contributes to the proper expression of gene products. Messenger RNAs (mRNAs) in eukaryotic cells possess a 5' cap structure and the 3' poly(A) tail which are important for mRNA stability and efficient translation. The Ccr4-Not complex is a major cytoplasmic deadenylase and functions in mRNA degradation. The CLB1-6 genes in Saccharomyces cerevisiae encode B-type cyclins which are involved in the cell cycle progression together with the cyclin-dependent kinase Cdc28. The CLB genes consist of CLB1/2, CLB3/4, and CLB5/6 whose gene products accumulate at the G2-M, S-G2, and late G1 phase, respectively. These Clb protein levels are thought to be mainly regulated by the transcriptional control and the protein stability control. Here we investigated regulation of CLB1-6 expression by Ccr4. Our results show that all CLB1-6 mRNA levels were significantly increased in the ccr4Δ mutant compared to those in wild-type cells. Clb1, Clb4, and Clb6 protein levels were slightly increased in the ccr4Δ mutant, but the Clb2, Clb3, and Clb5 protein levels were similar to those in wild-type cells. Since both CLB6 mRNA and Clb6 protein levels were most significantly increased in the ccr4Δ mutant, we further analyzed the cis-elements for the Ccr4-mediated regulation within CLB6 mRNA. We found that there were destabilizing sequences in both coding sequence and 3' untranslated region (3' UTR). The destabilizing sequences in the coding region were found to be both within and outside the sequences corresponding the cyclin domain. The CLB6 3' UTR was sufficient for mRNA destabilization and decrease of the reporter GFP gene and this destabilization involved Ccr4. Our results suggest that CLB6 expression is regulated by Ccr4 through the coding sequence and 3' UTR of CLB6 mRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / genetics
  • Cyclin B* / metabolism
  • Cyclins / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Ribonucleases* / genetics
  • Ribonucleases* / metabolism
  • S Phase
  • Saccharomyces cerevisiae Proteins* / genetics
  • Saccharomyces cerevisiae Proteins* / metabolism
  • Saccharomyces cerevisiae* / metabolism

Substances

  • 3' Untranslated Regions
  • CLB4 protein, S cerevisiae
  • CLB5 protein, S cerevisiae
  • CLB6 protein, S cerevisiae
  • Cyclin B
  • Cyclins
  • RNA, Messenger
  • Saccharomyces cerevisiae Proteins
  • CCR4 protein, S cerevisiae
  • Ribonucleases

Grants and funding

This research was supported by JSPS KAKENHI Grant Number 18K06053 (to KI).