The yeast MATa1 gene contains two introns.

Am, Miller

doi:10.1002/j.1460-2075.1984.tb01927.x

Cited by 133 publications

(74 citation statements)

References 18 publications

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“…Among nuclear genes in S. cerevisiae, only the actin gene (18), the MATal gene (38), and several genes encoding ribosomal proteins (rpSl [60], S10 [31], and cyh2 [25]) have been shown to contain intervening sequences. Analysis of additional genes from ascomycetes is necessary to establish whether differences in the frequency of intervening sequences exist between the single-celled and filamentous ascomycetes.…”

Section: Resultsmentioning

confidence: 99%

Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori.

Nunberg¹,

Meade²,

Cole³

et al. 1984

Mol. Cell. Biol.

160

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The filamentous ascomycete Aspergillus awamori secretes large amounts of glucoamylase upon growth in medium containing starch, glucose, or a variety of hexose sugars and sugar polymers. We examined the mechanism of this carbon source-dependent regulation of glucoamylase accumulation and found a several hundredfold increase in glucoamylase mRNA in cells grown on an inducing substrate, starch, relative to cells grown on a noninducing substrate, xylose. We postulate that induction of glucoamylase synthesis is regulated transcriptionally. Comparing total mRNA from cells grown on starch and xylose, we were able to identify an inducible 2.3-kilobase mRNA-encoding glucoamylase. The glucoamylase mRNA was purified and used to identify a molecularly cloned 3.4-kilobase EcoRI fragment containing the A. awamori glucoamylase gene. Comparison of the nucleotide sequence of the 3.4-kilobase EcoRI fragment with that of the glucoamylase I mRNA (as determined from molecularly cloned cDNA) revealed the existence of four intervening sequences within the glucoamylase gene. The 5' end of the glucoamylase mRNA was mapped to several locations within a region -52 to -73 nucleotides from the translational start. Sequence and structural features of the glucoamylase gene of the filamentous ascomycete A. awamori were examined and compared with those reported in genes of other eucaryotes.

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Section: Resultsmentioning

confidence: 99%

Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori.

Nunberg¹,

Meade²,

Cole³

et al. 1984

Mol. Cell. Biol.

160

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“…The MATal transcript levels serve as an internal control for the amount of total RNA analyzed at each time point. The MATal transcript is spliced, resulting in four different size classes of protected fragments (Miller 1984 (Fig. 3A).…”

Section: R E S U L T Smentioning

confidence: 99%

“…Uniformly labeled probes homologous to each message were made by primer extension of M13 single-stranded DNA. The HO, HO :lacZ, H2B (HTBI), MATal, and SIR3 probes have been described previously (Miller 1984;Miller et al 1985;Nasmyth 1985a, b;Breeden and Nasmyth 1987a). The SWI4 probe used in Figures 1 and 2 includes DNA from about -300 to + 600 (from the ATG) cloned into M13mpl8.…”

Section: Transcript Analysismentioning

confidence: 99%

Cell cycle-specific expression of the SWI4 transcription factor is required for the cell cycle regulation of HO transcription.

Breeden¹,

Mikesell²

1991

Genes Dev.

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Expression of the HO endonuclease triggers mating-type switching in Saccharomyces cerevisiae. Transcription of the HO gene is start-dependent and restricted to the late G1/early S phase of haploid mother cells. The HO promoter contains 10 copies of a cell cycle-regulated upstream activation sequence, which is activated by SWI4 and SWI6. SWI4 mRNA levels vary at least 10-fold throughout the cell cycle and rise sharply just before the rise in HO mRNA levels. Constitutive synthesis of SWI4 mRNA leads to constitutive synthesis of HO mRNA. These data suggest that the cell cycle regulation of SWI4 mRNA is required for the tight cell cycle regulation of HO transcription. High-level constitutive synthesis of SWI4 also suppresses swi5 and swi6 mutations, suggesting that SWI4 is the predominant activator of HO transcription and that mutations in negative regulators of SWI4 could be isolated as suppressors of swi6 mutations. One recessive suppressor of swi6 (ssxl-1) that allowed high-level expression of SWI4 during ~x-factor arrest and constitutive expression of both SWI4 and HO after release from the arrest was isolated. This result suggests that SSX1 has a negative regulatory role in the cell-cycle regulation of SWI4 mRNA accumulation.

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“…RNAs were prepared from strains containing both the rapt~alleles and either a wild-type, hmr&A, or h m r h B silencer after a shift from 25°C to 37°C. The steady-state level of the HMRal transcript was quantified by an S1 nuclease protection assay, using the SIR3 transcript as an internal control (Miller 1984). In strains containing the rap1-5 allele, this analysis detected al transcripts 4 hr after the temperature shift in a strain containing the h m r h A silencer and 8 hr after the shift in a strain contaming the h m r h B silencer (Fig.…”

Section: A Heterologous Promoter At M a T A Restores A-specific Functmentioning

confidence: 99%

“…RNAs were precipitated in ethanol and analyzed by Northern blotting or with S1 nuclease after hybridization with prime-cut probes for the MATe,1 and SIR3 transcripts, as described (Miller 1984). Northern blots were prepared after electrophoretic separation of 20 ~g of total RNA for each time point on 1.4% agarose-formaldehyde gels.…”

Section: Isolation and Analysis Of Rnamentioning

confidence: 99%

RAP1 protein activates and silences transcription of mating-type genes in yeast.

1991

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RAP1 is a sequence-specific DNA-binding protein essential for cell growth. The occurrence of RAPl-binding sites in many promoter regions, the mating-type gene silencer elements, and telomeres suggests that RAP1 has multiple functions in the cell. To assess its role in transcription, temperature-sensitive mutations in RAP1 were generated. Analysis of rapl ts strains provides evidence that RAP1 functions in both transcriptional activation and silencing of mating-type genes. Several observations indicate that rap1 ts strains are defective in the expression of MATa, whose upstream activation sequence (UAS) contains a RAPl-binding site. At nonpermissive temperatures, decreases in MATa steady-state transcript levels can be detected in MATa ropl ts strains. Furthermore, these strains are deficient in a-pheromone production and simultaneously express at least two a-specific genes. These phenotypes can be reversed by replacing the RAPl-binding site at MAlTa with a binding site for the GALA transcriptional activator. Certain rap1 t~ alleles have an opposite effect on the silent mating-type locus HMR, which becomes partially derepressed at nonpermissive temperatures.

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The yeast MATa1 gene contains two introns.

Cited by 133 publications

References 18 publications

Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori.

Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori.

Cell cycle-specific expression of the SWI4 transcription factor is required for the cell cycle regulation of HO transcription.

RAP1 protein activates and silences transcription of mating-type genes in yeast.

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