Phenotypes Associated with Saccharomyces Cerevisiae Hug1 Protein, a Putative Negative Regulator of DNTP Levels, Reveal Similarities and Differences with Sequence-related Dif1

Overview

Journal J Microbiol

Specialty Microbiology

Date 2011 Mar 4

PMID 21369983

Citations 3

Authors

Eunmi Kim

Wolfram Siede

Affiliations

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Abstract

Saccharomyces cerevisiae Hugl is a small protein of unknown function that is highly inducible following replication stress and DNA damage. Its deletion suppresses the lethality of deletion of checkpoint kinase Mecl. Although DNA damage responses were largely normal in the HUG1 deletion mutant, we found enhanced resistance towards heat in logarithmic phase. In response to simultaneous carbon and replication stress, overall growth delay and less pseudohyphal filament formation were evident. These novel phenotypes are shared with deletion mutants of the negative regulators of ribonucleotide reductase, Difl and Smll. Microarray analysis showed the influence of Hugl on the expression of a large number of transcripts, including stress-related transcripts. Elevated dNTP levels in hugl Δ cells may result in a stress response reflected by the observed phenotypes and transcript profiles. However, in contrast to a deletion of structurally related Difl, Rnr2-Rnr4 subcellular localization is not grossly altered in a Hugl deletion mutant. Thus, although Hugl appears to be derived from the Rnr2-Rnr4 binding region of Difl, its mechanism of action must be independent of determining the localization of Rnr2-Rnr4.

Citing Articles

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References

Huang M, Zhou Z, Elledge S . The DNA replication and damage checkpoint pathways induce transcription by inhibition of the Crt1 repressor. Cell. 1998; 94(5):595-605. DOI: 10.1016/s0092-8674(00)81601-3. View

Zhao X, Chabes A, Domkin V, Thelander L, Rothstein R . The ribonucleotide reductase inhibitor Sml1 is a new target of the Mec1/Rad53 kinase cascade during growth and in response to DNA damage. EMBO J. 2001; 20(13):3544-53. PMC: 125510. DOI: 10.1093/emboj/20.13.3544. View

Lazzaro F, Giannattasio M, Puddu F, Granata M, Pellicioli A, Plevani P . Checkpoint mechanisms at the intersection between DNA damage and repair. DNA Repair (Amst). 2009; 8(9):1055-67. DOI: 10.1016/j.dnarep.2009.04.022. View

Lee Y, Elledge S . Control of ribonucleotide reductase localization through an anchoring mechanism involving Wtm1. Genes Dev. 2006; 20(3):334-44. PMC: 1361704. DOI: 10.1101/gad.1380506. View

Mizukami-Murata S, Iwahashi H, Kimura S, Nojima K, Sakurai Y, Saitou T . Genome-wide expression changes in Saccharomyces cerevisiae in response to high-LET ionizing radiation. Appl Biochem Biotechnol. 2010; 162(3):855-70. DOI: 10.1007/s12010-009-8825-3. View

Zhao X, Rothstein R . The Dun1 checkpoint kinase phosphorylates and regulates the ribonucleotide reductase inhibitor Sml1. Proc Natl Acad Sci U S A. 2002; 99(6):3746-51. PMC: 122595. DOI: 10.1073/pnas.062502299. View

Chabes A, Georgieva B, Domkin V, Zhao X, Rothstein R, Thelander L . Survival of DNA damage in yeast directly depends on increased dNTP levels allowed by relaxed feedback inhibition of ribonucleotide reductase. Cell. 2003; 112(3):391-401. DOI: 10.1016/s0092-8674(03)00075-8. View

Jiang Y, Kang C . Induction of S. cerevisiae filamentous differentiation by slowed DNA synthesis involves Mec1, Rad53 and Swe1 checkpoint proteins. Mol Biol Cell. 2003; 14(12):5116-24. PMC: 284813. DOI: 10.1091/mbc.e03-06-0375. View

Yao R, Zhang Z, An X, Bucci B, Perlstein D, Stubbe J . Subcellular localization of yeast ribonucleotide reductase regulated by the DNA replication and damage checkpoint pathways. Proc Natl Acad Sci U S A. 2003; 100(11):6628-33. PMC: 164498. DOI: 10.1073/pnas.1131932100. View

10.

Benton M, Glasser N, Palecek S . The utilization of a Saccharomyces cerevisiae HUG1P-GFP promoter-reporter construct for the selective detection of DNA damage. Mutat Res. 2007; 633(1):21-34. DOI: 10.1016/j.mrgentox.2007.05.002. View

11.

Nyberg K, Michelson R, Putnam C, Weinert T . Toward maintaining the genome: DNA damage and replication checkpoints. Annu Rev Genet. 2002; 36:617-56. DOI: 10.1146/annurev.genet.36.060402.113540. View

12.

Kastan M, Bartek J . Cell-cycle checkpoints and cancer. Nature. 2004; 432(7015):316-23. DOI: 10.1038/nature03097. View

13.

Pabla R, Pawar V, Zhang H, Siede W . Characterization of checkpoint responses to DNA damage in Saccharomyces cerevisiae: basic protocols. Methods Enzymol. 2006; 409:101-17. DOI: 10.1016/S0076-6879(05)09006-3. View

14.

Huang D, Friesen H, Andrews B . Pho85, a multifunctional cyclin-dependent protein kinase in budding yeast. Mol Microbiol. 2007; 66(2):303-14. DOI: 10.1111/j.1365-2958.2007.05914.x. View

15.

Wu X, Huang M . Dif1 controls subcellular localization of ribonucleotide reductase by mediating nuclear import of the R2 subunit. Mol Cell Biol. 2008; 28(23):7156-67. PMC: 2593381. DOI: 10.1128/MCB.01388-08. View

16.

Lee Y, Wang J, Stubbe J, Elledge S . Dif1 is a DNA-damage-regulated facilitator of nuclear import for ribonucleotide reductase. Mol Cell. 2008; 32(1):70-80. PMC: 3245869. DOI: 10.1016/j.molcel.2008.08.018. View

17.

Nestoras K, Mohammed A, Schreurs A, Fleck O, Watson A, Poitelea M . Regulation of ribonucleotide reductase by Spd1 involves multiple mechanisms. Genes Dev. 2010; 24(11):1145-59. PMC: 2878652. DOI: 10.1101/gad.561910. View

18.

Basrai M, Velculescu V, Kinzler K, Hieter P . NORF5/HUG1 is a component of the MEC1-mediated checkpoint response to DNA damage and replication arrest in Saccharomyces cerevisiae. Mol Cell Biol. 1999; 19(10):7041-9. PMC: 84699. DOI: 10.1128/MCB.19.10.7041. View

19.

Rothstein R . Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. Methods Enzymol. 1991; 194:281-301. DOI: 10.1016/0076-6879(91)94022-5. View

20.

Zaim J, Speina E, Kierzek A . Identification of new genes regulated by the Crt1 transcription factor, an effector of the DNA damage checkpoint pathway in Saccharomyces cerevisiae. J Biol Chem. 2004; 280(1):28-37. DOI: 10.1074/jbc.M404669200. View