Two differentially regulated mRNAs with different 5′ ends encode secreted and intracellular forms of yeast invertase

Carlson, Marian; Botstein, David

doi:10.1016/0092-8674(82)90384-1

Cited by 1,538 publications

(860 citation statements)

References 33 publications

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“…Total RNA was isolated according to Carlson and Botstein (1982) and Northern blot analysis was performed as described by Má rquez et al (1998). The probes used were PCR fragments spanning the whole CTT1 and HAL2 ORFs or part of the NST1 ORF.…”

Section: Northern Blot Analysismentioning

confidence: 99%

Involvement of Nst1p/YNL091w and Msl1p, a U2B″ splicing factor, in Saccharomyces cerevisiae salt tolerance

Goossens¹,

Forment²,

Serrano³

2002

Yeast

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Cell tolerance to salt stress depends on many physiological functions, including the best characterized of osmotic adjustment, ion transport and sodium-sensitive sulphate metabolism. From a screening designed to identify novel determinants of salt tolerance we have isolated the YNL091w gene, probably an Ascomycete-specific gene encoding a protein of unknown function. This gene negatively affects salt tolerance and therefore has been designated NST1. The salt tolerance mechanism of nst1 mutants is novel because it is not related to osmoregulation, altered cation accumulation or sulphate metabolism. Genome-wide two-hybrid analysis has suggested that Nst1p interacts with the splicing factor Msl1p and, accordingly, the impact of NST1 on salt tolerance is dependent on a functional MSL1 gene. Loss of MSL1 and NST1 function has pleiotropic phenotypes including increased sensitivity to divalent cations (manganese and zinc) and to caffeine (a cell wall-weakening agent). On the other hand, msl1 mutants but not nst1 mutants are sensitive to thiabendazole (a microtubule-destabilizing agent) and to osmotic stress.

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Section: Northern Blot Analysismentioning

confidence: 99%

Involvement of Nst1p/YNL091w and Msl1p, a U2B″ splicing factor, in Saccharomyces cerevisiae salt tolerance

Goossens¹,

Forment²,

Serrano³

2002

Yeast

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“…A genomic library carried by a 2 micron (2 mm) plasmid vector (Yep24; Carlson & Botstein 1982) was introduced into a dmc1D/dmc1D diploid strain by selection for uracil prototrophy. Following growth on medium that selected for the maintenance of plasmids (ÀUra medium), individual transformed clones were transferred as patches on to a nitrocellulose ®lter that had been placed on the surface of a ÀUra plate.…”

Section: Screen For High Copy Suppressors Of Dmc1 Arrestmentioning

confidence: 99%

“…This mutation allows maintenance of a cloned sequence at two different copy numbers levels depending on the growth conditions; selection for Ura causes YEpFAT4 to be maintained at an average of approximately 20 copies per cell (a level typical for standard YEp 2m plasmids) and selection for Leu results in selection of cells with an average of over 100 copies per cell (K. Runge, personal communication). YepFAT4-RAD54 (pNRB143) was constructed by ®rst inserting the 3.5kb RAD54-containing NheI fragment (SGD coordinates VII-193477-197033) from the original library isolate into pRS426 (Christianson et al 1992) and then transferring the insert as a SmaI-SacII fragment into SmaI-SacI cut YepFAT4. A derivative of this plasmid containing a frame shift mutation in RAD54 (pNRB233) was generated by ®lling in the AvrII site of pNRB143 using the Klenow fragment of E. coli DNA polymerase I. YepFAT4-REC114 (pNRB235) was constructed by inserting the REC114-containing 2.4 kB AvrII-NruI kb fragment (SGD coordinates XIII-535734-538159) from the library isolate into XbaI-SmaI cut YepFAT4.…”

Section: Plasmidsmentioning

confidence: 99%

High copy number suppression of the meiotic arrest caused by a dmc1 mutation: REC114 imposes an early recombination block and RAD54 promotes a DMC1‐independent DSB repair pathway

et al. 1999

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Background: DMC1, the meiosis-speci®c eukaryotic homologue of bacterial recA, is required for completion of meiotic recombination and cell cycle progression past prophase. In a dmc1 mutant, double strand break recombination intermediates accumulate and cells arrest in prophase. We isolated genes which, when present at high copy numbers, suppress the meiotic arrest phenotype conferred by dmc1 mutations.

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“…This enzyme, which hydrolyzes sucrose, is encoded by the yeast SUC2 gene. A cytoplasmic form of invertase is constitutively expressed, whereas the secreted form of invertase is produced only in response to glucose limitation (Carlson and Botstein, 1982). The secreted form of invertase is core glycosylated in the endoplasmic reticulum and further glycosylated in the Golgi apparatus.…”

Section: Phenotypes Of Arf1 Mutantsmentioning

confidence: 99%

Systematic Structure-Function Analysis of the Small GTPase Arf1 in Yeast

Click¹,

Stearns

Botstein³

2002

MBoC

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Members of the ADP-ribosylation factor (Arf) family of small GTPases are implicated in vesicle traffic in the secretory pathway, although their precise function remains unclear. We generated a series of 23 clustered charge-to-alanine mutations in the Arf1 protein ofSaccharomyces cerevisiae to determine the portions of this protein important for its function in cells. These mutants display a number of phenotypes, including conditional lethality at high or low temperature, defects in glycosylation of invertase, dominant lethality, fluoride sensitivity, and synthetic lethality with thearf2 null mutation. All mutations were mapped onto the available crystal structures for Arf1p: Arf1p bound to GDP, to GTP, and complexed with the regulatory proteins ArfGEF and ArfGAP. From this systematic structure-function analysis we demonstrate that all essential mutations studied map to one hemisphere of the protein and provide strong evidence in support of the proposed ArfGEF contact site on Arf1p but minimal evidence in support of the proposed ArfGAP-binding site. In addition, we describe the isolation of a spatially distant intragenic suppressor of a dominant lethal mutation in the guanine nucleotide-binding region of Arf1p.

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Two differentially regulated mRNAs with different 5′ ends encode secreted and intracellular forms of yeast invertase

Cited by 1,538 publications

References 33 publications

Involvement of Nst1p/YNL091w and Msl1p, a U2B″ splicing factor, in Saccharomyces cerevisiae salt tolerance

Involvement of Nst1p/YNL091w and Msl1p, a U2B″ splicing factor, in Saccharomyces cerevisiae salt tolerance

High copy number suppression of the meiotic arrest caused by a dmc1 mutation: REC114 imposes an early recombination block and RAD54 promotes a DMC1‐independent DSB repair pathway

Systematic Structure-Function Analysis of the Small GTPase Arf1 in Yeast

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