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  • 1
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; cAMP-dependent protein kinase ; Regulatory subunit ; Site-directed mutagenesis ; Phosphorylation site
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Four mutants with amino acid substitution(s) at or near the putative phosphorylation site (Arg142 Arg143 Thr144 Ser145) of the regulatory subunit of cAMP-dependent protein kinase were obtained by site-directed mutagenesis. Three mutants, BCY1 Ala 145 (Ser145 to Ala), BCY1 His 143 (Arg143 to His) and BCY1 Asn 144, Ala 145 (Thr144 to Asn and Ser145 to Ala) complemented a bcy1 mutant, whereas BCY1 Gly 143 (Arg143 to Gly) did not. In addition, mutant, BCY1 Asn 144, Ala 145 exhibited a dominant coldsensitive phenotype, which can be most easily explained by the functional alteration of the regulatory subunit of cAMP-dependent protein kinase by the mutations. Analyses of these mutant genes revealed that phosphorylation of the regulatory subunit is not a prerequisite for the regulation of the cAMP-dependent protein kinase activity in responding to the cAMP level.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: G1 cyclin ; RNA binding protein ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three G1 cyclins,CLN1,CLN2, andCLN3, have been identified in the budding yeastSaccharomyces cerevisiae. G1 cyclins are essential, albeit functionally redundant, rate-limiting activators of cell cycle initiation. We have isolated dosage-dependent suppressor genes (designatedHMD genes) of the mating defect caused byCLN3-2, a dominant mutation inCLN3,HMD2 andHMD3 are identical toSTE4 andSTE5, respectively,HMD1 is an essential gene that encodes a protein containing a putative RNA binding domain. Overproduction ofHMD1 results in a relatively specific reduction in the level of theCLN3 orCLN3-2 transcript. This reduction occurs subsequent to transcription initiation ofCLN3 since overexpression ofHMD1 did not affect expression of a heterologous transcript from theCLN3 promoter but did result in a reduction ofCLN3 transcript expressed from a heterologous promoter.HMD1 has at least one essential role independent of its effect onCLN3 sinceHMD1 remains essential for viability in the absence of a functionalCLN3 gene.
    Type of Medium: Electronic Resource
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