Disruption of six novelSaccharomyces cerevisiae genes reveals thatYGL129c is necessary for growth in non-fermentable carbon sources,YGL128c for growth at low or high temperatures andYGL125w is implicated in the biosynthesis of methionine

Tizon, Belen; Rodríguez-Torres, Ana María; Cerdán, M. Esperanza

doi:10.1002/(sici)1097-0061(19990130)15:2<145::aid-yea346>3.0.co;2-j

Cited by 24 publications

(15 citation statements)

References 24 publications

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“…Alternatively reported as essential (Giaever et al 2002) or nonessential (Tizon et al 1999), we find the cwc23TKAN null mutant to be viable but sickly. This stain is also genetically unstable, giving rise to more rapidly growing suppressors, an observation perhaps relevant to the diversity of cwc23 phenotypes reported (S. Pandit, M. Chen and B. C. Rymond, unpublished results).…”

Section: Discussionmentioning

confidence: 44%

“…This motif is used by bacterial DnaJ (or eukaryotic Hsp40) proteins to activate the ATPase activity of a companion Hsp70 protein during macromolecular assembly and disassembly events (Walsh et al 2004;Vos et al 2008). Null mutants of CWC23 were previously described as lethal (Giaever et al 2002) or viable but temperature and cold sensitive (Tizon et al 1999). We were able to isolate slow-growing yeast without CWC23 by 5-fluorootic acid (5-FOA) selection against a plasmid-borne wild-type allele in a cwc23TKAN chromosomal background lacking the entire CWC23 protein coding sequence ( Figure 6A).…”

Section: Resultsmentioning

confidence: 99%

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Spp382p Interacts with Multiple Yeast Splicing Factors, Including Possible Regulators of Prp43 DExD/H-Box Protein Function

Pandit

Paul

Zhang³

et al. 2009

Genetics

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Prp43p catalyzes essential steps in pre-mRNA splicing and rRNA biogenesis. In splicing, Spp382p stimulates the Prp43p helicase to dissociate the postcatalytic spliceosome and, in some way, to maintain the integrity of the spliceosome assembly. Here we present a dosage interference assay to identify Spp382p-interacting factors by screening for genes that when overexpressed specifically inhibit the growth of a conditional lethal prp38-1 spliceosome assembly mutant in the spp382-1 suppressor background. Identified, among others, are genes encoding the established splicing factors Prp8p, Prp9p, Prp11p, Prp39p, and Yhc1p and two poorly characterized proteins with possible links to splicing, Sqs1p and Cwc23p. Sqs1p copurifies with Prp43p and is shown to bind Prp43p and Spp382p in the two-hybrid assay. Overexpression of Sqs1p blocks pre-mRNA splicing and inhibits Prp43p-dependent steps in rRNA processing. Increased Prp43p levels buffer Sqs1p cytotoxicity, providing strong evidence that the Prp43p DExD/H-box protein is a target of Sqs1p. Cwc23p is the only known yeast splicing factor with a DnaJ motif characteristic of Hsp40-like chaperones. We show that similar to SPP382, CWC23 activity is critical for efficient pre-mRNA splicing and intron metabolism yet, surprisingly, this activity does not require the canonical DnaJ/Hsp40 motif. These and related data establish the value of this dosage interference assay for finding genes that alter cellular splicing and define Sqs1p and Cwc23p as prospective modulators of Spp382p-stimuated Prp43p function. E IGHT phylogenetically conserved DExD/H-box proteins act at discrete steps to regulate the assembly, activation, and dissociation of the splicing apparatus (reviewed in Brow 2002; Konarska and Query 2005;Linder 2006). How these RNA-dependent ATPases are temporally and functionally regulated remains poorly understood. One putative regulator, the 83-kDa Spp382/Ntr1 protein (henceforth referred to by the Saccharomyces Genome Database standard name, Spp382p), was discovered in a screen for mutants capable of suppressing defects in yeast spliceosome assembly (Pandit et al. 2006). While spp382 null alleles are lethal, partial loss of function suppresses mutations in several other splicing factors, including the genes encoding the essential spliceosomal proteins Prp8p and Prp38p. Spp382p is a spliceosomal protein that binds the Prp43p DExD/H-box protein to promote efficient dissociation of spliceosomal factors after completion of splicing in vitro (Tsai et al. 2005). Some but not all spp382 mutants also accumulate the excised intron product of splicing in vivo, ostensibly due to protection of the intron within a hyperstabilized spliceosome (Pandit et al. 2006;Tanaka et al. 2007). The suppression of spliceosome assembly defects by spp382 mutation is proposed to occur by impairing Spp382p-stimulated dissociation of kinetically impaired or otherwise inefficient spliceosomes by Prp43p (Pandit et al. 2006). Consistent with this hypothesis, prp43 mutations also suppress spliceosome a...

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Section: Discussionmentioning

confidence: 44%

Section: Resultsmentioning

confidence: 99%

Spp382p Interacts with Multiple Yeast Splicing Factors, Including Possible Regulators of Prp43 DExD/H-Box Protein Function

Pandit

Paul

Zhang³

et al. 2009

Genetics

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“…is encoded by the MET13 gene product that supplies onecarbon units for the synthesis of methionine (28). This pathway transfers the methyl moiety via S-adenosylmethionine to the C-4 position of choline (resonating at ϳ55.3 ppm in 13 C NMR spectra; Table III).…”

Section: Resultsmentioning

confidence: 99%

Regulation of the Balance of One-carbon Metabolism inSaccharomyces cerevisiae

Piper¹,

Hong²,

Ball³

et al. 2000

Journal of Biological Chemistry

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One-carbon metabolism in yeast is an essential process that relies on at least one of three one-carbon donor molecules: serine, glycine, or formate. By a combination of genetics and biochemistry we have shown how cells regulate the balance of one-carbon flow between the donors by regulating cytoplasmic serine hydroxymethyltransferase activity in a side reaction occurring in the presence of excess glycine. This control governs the level of 5,10-methylene tetrahydrofolate (5,10-CH 2 -H 4 folate) in the cytoplasm, which has a direct role in signaling transcriptional control of the expression of key genes, particularly those encoding the unique components of the glycine decarboxylase complex (GCV1, GCV2, and GCV3). Based on these and other observations, we propose a model for how cells balance the need to supplement their one-carbon pools when charged folates are limiting or when glycine is in excess. We also propose that under normal conditions, cytoplasmic 5,10-CH 2 -H 4 folate is mainly directed to generating methyl groups via methionine, whereas one-carbon units generated from glycine in mitochondria are more directed to purine biosynthesis. When glycine is in excess, 5,10-CH 2 -H 4 folate is decreased, and the regulation loop shifts the balance of generation of one-carbon units into the mitochondrion.

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“…The correct replacement in the S. cerevisiae g e n o m e w a s v e r i f i e d b y P C R a s previously described (Tizón et al, 1999). Genomic DNAs isolated from the BY4741-∆ixr1 and the null candidates were amplified with two pairs of primers.…”

Section: Yeast Strains and Construction Of Double Knock-outsmentioning

confidence: 99%

The Yeast Genes ROX1, IXR1, SKY1 and Their Effect upon Enzymatic Activities Related to Oxidative Stress

Leiro¹,

Lombardero²,

Vázquez³

et al. 2012

Oxidative Stress - Molecular Mechanisms and Biological Effects

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Disruption of six novelSaccharomyces cerevisiae genes reveals thatYGL129c is necessary for growth in non-fermentable carbon sources,YGL128c for growth at low or high temperatures andYGL125w is implicated in the biosynthesis of methionine

Cited by 24 publications

References 24 publications

Spp382p Interacts with Multiple Yeast Splicing Factors, Including Possible Regulators of Prp43 DExD/H-Box Protein Function

Spp382p Interacts with Multiple Yeast Splicing Factors, Including Possible Regulators of Prp43 DExD/H-Box Protein Function

Regulation of the Balance of One-carbon Metabolism inSaccharomyces cerevisiae

The Yeast Genes ROX1, IXR1, SKY1 and Their Effect upon Enzymatic Activities Related to Oxidative Stress

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