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Molecular and Cellular Biology, January 2002, p. 1-11, Vol. 22, No. 1
0270-7306/01/$04.00+0     DOI: 10.1128/MCB.22.1.1-11.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Protein Kinase A Regulates Sexual Development and Gluconeogenesis through Phosphorylation of the Zn Finger Transcriptional Activator Rst2p in Fission Yeast

Toru Higuchi, Yoshinori Watanabe, and Masayuki Yamamoto*

Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Hongo, Tokyo 113-0033, Japan

Received 12 June 2001/ Returned for modification 26 July 2001/ Accepted 26 September 2001

Protein kinase A (PKAi a cyclic AMP-dependent protein kinase) negatively regulates sexual development and gluconeogenesis in fission yeast by suppressing the transcription of ste11 required for the former and the transcription of fbp1 required for the latter. Here we show that Rst2p, a zinc finger protein that can bind to the upstream region of ste11 and fbp1 via the STREP motif, mediates the activity of PKA to transcription of these genes. A simple reporter system confirmed that PKA could cause its negative effect on transcription through the combination of Rst2p and STREP. Rst2p was phosphorylated by PKA in vitro at two consensus sequences on it. Substitution of the target threonine residues by alanine made the protein active even in the presence of high PKA activity. Rst2p underwent hyperphosphorylation in the medium lacking glucose, and PKA inhibited this hyperphosphorylation. Rst2p was mainly cytoplasmic under high PKA activity but was concentrated in the nucleus when this activity was lowered, suggesting that PKA might regulate ste11 and fbp1 negatively by excluding Rst2p from the nucleus. However, the shift of Rst2p localization was not perfect under physiological conditions, leaving the possibility that PKA inhibits Rst2p function in another way as well. Although the PKA-Rst2p-STREP pathway is apparently central to the regulation of ste11 and fbp1 transcription in accordance with nutritional conditions, some additional paths are likely to connect nitrogen to repression of ste11 and glucose to repression of fbp1. These paths may ensure the specificity between the type of nutrients in shortage and the type of genes to be expressed.


* Corresponding author. Mailing address: Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Hongo, Tokyo 113-0033, Japan. Phone: 81-3-3814-9620. Fax: 81-3-5802-2042. E-mail address: myamamot{at}ims.u-tokyo.ac.jp.


Molecular and Cellular Biology, January 2002, p. 1-11, Vol. 22, No. 1
0022-538X/01/$04.00+0     DOI: 10.1128/MCB.22.1.1-11.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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