Characterization of sua7 mutations defines a domain of TFIIB involved in transcription start site selection in yeast

Inés Pinto, Wei Hua Wu, Jong G. Na, Michael Hampsey

    Research output: Contribution to journalArticle

    100 Citations (Scopus)

    Abstract

    The SUA7 gene of Saccharomyces cerevisiae encodes the general transcription factor TFIIB. SUA7 was identified based on the ability of mutations at this locus to shift transcription start site selection at the cyc1 gene downstream of normal. Here, we report the nature of these mutations; the sua7-1 and sua7-2 alleles encode identical E62K replacements, and sua7-3 encodes an R78C replacement. Both Glu-62 and Arg-78 are phylogenetically invariant and occur within the most highly conserved region of TFIIB, immediately distal to a zinc finger motif. A double E62K,R78C mutant was constructed and exhibited the same phenotypes associated with the single mutants, including cold sensitivity and altered start site selection, suggesting that Glu-62 and Arg-78 are functionally related. This observation, and the opposite charge of the 2 residues, suggested that Glu-62 and Arg-78 might interact to form a salt bridge. This was tested by constructing reciprocal E62R and R78E replacements. The E62R mutant is phenotypically identical to the E62K mutant, whereas the R78E mutant is inviable. However, an E62R,R78E double mutant was not only viable but is phenotypically similar to the single mutants. These results define the highly conserved sequence adjacent to the zinc finger of TFIIB as a critical determinant of start site selection and suggest that an Glu-62-Arg-78 salt bridge is an important structural element of that domain.

    Original languageEnglish (US)
    Pages (from-to)30569-30573
    Number of pages5
    JournalJournal of Biological Chemistry
    Volume269
    Issue number48
    StatePublished - Dec 2 1994

    Fingerprint

    Transcription Factor TFIIB
    Site selection
    Transcription Initiation Site
    Yeast
    Yeasts
    Zinc Fingers
    Mutation
    Zinc
    Salts
    Genes
    General Transcription Factors
    Conserved Sequence
    Saccharomyces cerevisiae
    Alleles
    Phenotype

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Characterization of sua7 mutations defines a domain of TFIIB involved in transcription start site selection in yeast. / Pinto, Inés; Wu, Wei Hua; Na, Jong G.; Hampsey, Michael.

    In: Journal of Biological Chemistry, Vol. 269, No. 48, 02.12.1994, p. 30569-30573.

    Research output: Contribution to journalArticle

    Pinto, Inés ; Wu, Wei Hua ; Na, Jong G. ; Hampsey, Michael. / Characterization of sua7 mutations defines a domain of TFIIB involved in transcription start site selection in yeast. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 48. pp. 30569-30573.
    @article{341357e6cfd142a8831f75435c85a7f7,
    title = "Characterization of sua7 mutations defines a domain of TFIIB involved in transcription start site selection in yeast",
    abstract = "The SUA7 gene of Saccharomyces cerevisiae encodes the general transcription factor TFIIB. SUA7 was identified based on the ability of mutations at this locus to shift transcription start site selection at the cyc1 gene downstream of normal. Here, we report the nature of these mutations; the sua7-1 and sua7-2 alleles encode identical E62K replacements, and sua7-3 encodes an R78C replacement. Both Glu-62 and Arg-78 are phylogenetically invariant and occur within the most highly conserved region of TFIIB, immediately distal to a zinc finger motif. A double E62K,R78C mutant was constructed and exhibited the same phenotypes associated with the single mutants, including cold sensitivity and altered start site selection, suggesting that Glu-62 and Arg-78 are functionally related. This observation, and the opposite charge of the 2 residues, suggested that Glu-62 and Arg-78 might interact to form a salt bridge. This was tested by constructing reciprocal E62R and R78E replacements. The E62R mutant is phenotypically identical to the E62K mutant, whereas the R78E mutant is inviable. However, an E62R,R78E double mutant was not only viable but is phenotypically similar to the single mutants. These results define the highly conserved sequence adjacent to the zinc finger of TFIIB as a critical determinant of start site selection and suggest that an Glu-62-Arg-78 salt bridge is an important structural element of that domain.",
    author = "In{\'e}s Pinto and Wu, {Wei Hua} and Na, {Jong G.} and Michael Hampsey",
    year = "1994",
    month = "12",
    day = "2",
    language = "English (US)",
    volume = "269",
    pages = "30569--30573",
    journal = "Journal of Biological Chemistry",
    issn = "0021-9258",
    publisher = "American Society for Biochemistry and Molecular Biology Inc.",
    number = "48",

    }

    TY - JOUR

    T1 - Characterization of sua7 mutations defines a domain of TFIIB involved in transcription start site selection in yeast

    AU - Pinto, Inés

    AU - Wu, Wei Hua

    AU - Na, Jong G.

    AU - Hampsey, Michael

    PY - 1994/12/2

    Y1 - 1994/12/2

    N2 - The SUA7 gene of Saccharomyces cerevisiae encodes the general transcription factor TFIIB. SUA7 was identified based on the ability of mutations at this locus to shift transcription start site selection at the cyc1 gene downstream of normal. Here, we report the nature of these mutations; the sua7-1 and sua7-2 alleles encode identical E62K replacements, and sua7-3 encodes an R78C replacement. Both Glu-62 and Arg-78 are phylogenetically invariant and occur within the most highly conserved region of TFIIB, immediately distal to a zinc finger motif. A double E62K,R78C mutant was constructed and exhibited the same phenotypes associated with the single mutants, including cold sensitivity and altered start site selection, suggesting that Glu-62 and Arg-78 are functionally related. This observation, and the opposite charge of the 2 residues, suggested that Glu-62 and Arg-78 might interact to form a salt bridge. This was tested by constructing reciprocal E62R and R78E replacements. The E62R mutant is phenotypically identical to the E62K mutant, whereas the R78E mutant is inviable. However, an E62R,R78E double mutant was not only viable but is phenotypically similar to the single mutants. These results define the highly conserved sequence adjacent to the zinc finger of TFIIB as a critical determinant of start site selection and suggest that an Glu-62-Arg-78 salt bridge is an important structural element of that domain.

    AB - The SUA7 gene of Saccharomyces cerevisiae encodes the general transcription factor TFIIB. SUA7 was identified based on the ability of mutations at this locus to shift transcription start site selection at the cyc1 gene downstream of normal. Here, we report the nature of these mutations; the sua7-1 and sua7-2 alleles encode identical E62K replacements, and sua7-3 encodes an R78C replacement. Both Glu-62 and Arg-78 are phylogenetically invariant and occur within the most highly conserved region of TFIIB, immediately distal to a zinc finger motif. A double E62K,R78C mutant was constructed and exhibited the same phenotypes associated with the single mutants, including cold sensitivity and altered start site selection, suggesting that Glu-62 and Arg-78 are functionally related. This observation, and the opposite charge of the 2 residues, suggested that Glu-62 and Arg-78 might interact to form a salt bridge. This was tested by constructing reciprocal E62R and R78E replacements. The E62R mutant is phenotypically identical to the E62K mutant, whereas the R78E mutant is inviable. However, an E62R,R78E double mutant was not only viable but is phenotypically similar to the single mutants. These results define the highly conserved sequence adjacent to the zinc finger of TFIIB as a critical determinant of start site selection and suggest that an Glu-62-Arg-78 salt bridge is an important structural element of that domain.

    UR - http://www.scopus.com/inward/record.url?scp=0028019848&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0028019848&partnerID=8YFLogxK

    M3 - Article

    C2 - 7982976

    AN - SCOPUS:0028019848

    VL - 269

    SP - 30569

    EP - 30573

    JO - Journal of Biological Chemistry

    JF - Journal of Biological Chemistry

    SN - 0021-9258

    IS - 48

    ER -