Abstract
A central problem in our understanding of mitochondrial (mt) function remains the question of how coordinate transcriptional control is accomplished between nucleus and mitochondria. Here, we report the initial characterization of a protein of previously unknown function, the product of the YMR030 W gene, that appears to mediate such coordinate gene expression. Expression of YMR030 W is glucose-repressible; a deletion mutant for this gene shows a severe growth defect on glycerol-, but not glucose- or ethanol-based medium. In that mutant, transcript levels from GUT1 and GUT2 are highly attenuated compared with those of the wild-type parent when both are grown on glycerol-based medium. Under the same growth conditions, transcripts from the mt OLI1 gene, which has one copy of a mt upstream activating sequence (UAS) in its 5′-flanking region, are attenuated in the ΔYMR030 W mutant, but mRNA from the mt COX3 (OXI2) gene, which lacks the mt UAS, are not. Some nuclear genes encoding mt-related proteins also show low transcript levels in the ΔYMR030 W mutant in comparison with those of the wild-type parent strain during glycerol-based growth. Localization of the protein, via its expression fused to green fluorescent protein, indicates that it is present in both nucleus and mitochondria, supporting a respiration-related transcriptional role for this gene product in both cellular genetic compartments. Because of its role in both respiratory growth and mt function, we designate the YMR030 W coding sequence RSF1 (respiration factor 1).
Similar content being viewed by others
References
Gérard HC, Krauße-Opatz B, Rudy D, Rao JP, Zeidler H, Schumacher HR, Whittum-Hudson JA, Köhler L, Hudson AP (2001) Expression of Chlamydia trachomatis genes required for DNA synthesis and cell division in active vs persistent infection. Mol Microbiol 41:731–741
Grauslund M, Rønnow B (2000) Carbon source-dependent transcriptional regulation of the mitochondrial glycerol-3-phosphate dehydrogenase gene, GUT2, from Saccharomyces cerevisiae. Can J Microbiol 46:1096–1100
Grauslund M, Lopes JM, Rønnow B (1999) Expression of GUT1, which encodes glycerol kinase in Saccharomyces cerevisiae, is controlled by the positive regulators Adr1p, Ino2p and Ino4p and the negative regulator Opi1; in a carbon source-dependent fashion. Nucleic Acids Res 27:4391–4398
Hiratsuka M, Agatsuma Y, Mizugaki M (1999) Rapid detection of CYP2C9*3 alleles by real time fluorescence PCR based on SYBR green. Mol Genet Metab 68:357–362
Iqbal J, Rahman MU, Gérard HC, Yu J, Nevel CA, Hudson AP (1996) A probable cis-regulatory element on yeast mitochondrial DNA responsible for cAMP-dependent transcription. Curr Genet 30:493–501
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
James P, Halladay J, Craig EA (1996) Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics 144:1425–1436
McEntee CM, Hudson AP (1989) Preparation of RNA from unspheroplasted yeast cells (Saccharomyces cerevisiae). Anal Biochem 176:303–306
McEntee CM, Cantwell R, Rahman MU, Hudson AP (1993) Transcription of the yeast mitochondrial genome requires cAMP. Mol Gen Genet 241:213–224
Pavlik P, Simon M, Schuster T, Ruis H (1993) The glycerol kinase (GUT1) gene of Saccharomyces cerevisiae: cloning and characterization. Curr Genet 24:21–25
Pfeifer K, Prezant T, Guarente L (1987) Yeast HAP1 activator binds to two upstream activation elements of different sequence. Cell 49:19–27
Rahman MU, Hudson AP (1995a) Nature and transcriptional role of catalytic subunits of yeast mitochondrial cAMP-dependent protein kinase (Saccharomyces cerevisiae). Biochem Biophys Res Commun 206:756–763
Rahman MU, Hudson AP (1995b) Substrates for yeast mitochondrial cAMP-dependent protein kinase activity. Biochem Biophys Res Commun 214:188–194
Rahman MU, Kleyman TR, McEntee CM, Hudson AP (1994) Regulation of mitochondrial cAMP-dependent protein kinase activity in yeast. Biochem Mol Biol Int 34:745–753
Rønnow B, Kielland-Brandt MC (1993) GUT2, a gene for mitochondrial glycerol-3-phosphate dehydrogenase of Saccharomyces cerevisiae. Yeast 9:1121–1130
Sherman F (1991) Getting started with yeast. In: Guthrie C, Fink GR (eds) Guide to yeast genetics and molecular biology. Methods Enzymol 194:3–21
Uetz, P, Giot L, Cagney G, Mansfield TA, Judson RS, Knight JR, Lockshon D, Narayan V, Srinivasan M, Pochart P, Qureshi-Emili A, Li Y, Godwin B, Conover D, Kalbfleisch T, Vihayadamodar G, Yang M, Johnston M, Fields S, Rothberg JM (2000) A comprehensive analysis of protein–protein interactions in Saccharomyces cerevisiae. Nature 403:623–627
Wang Z, Wu X, Levin RM, Hudson AP (2001) Loss of mitochondrial DNA in rabbit bladder smooth muscle following partial outlet obstruction results from lack of organellar DNA replication. Mol Urol 5:99–104
Acknowledgements
This work was supported by a Merit Review grant to A.P.H. from the Department of Veterans Affairs Medical Research Service and by a REAP Award from the same source to A.P.H. and other investigators at the Detroit DVA Medical Center. We are grateful to Dr. P. James for supplying the pGAD library used in the one-hybrid screening procedure and to Drs. L. Bergman, N. Davis, and J. Hegemann for supplying plasmids used in several experiments described here. We are grateful to Dr. J.A. Whittum-Hudson for use of her microscope.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by S. Hohmann
Rights and permissions
About this article
Cite this article
Lu, L., Roberts, G., Simon, K. et al. Rsf1p, a protein required for respiratory growth of Saccharomyces cerevisiae . Curr Genet 43, 263–272 (2003). https://doi.org/10.1007/s00294-003-0398-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00294-003-0398-z