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    Alanine-scanning mutagenesis of Aspergillus γ-tubulin yields diverse and novel phenotypes

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    Oakley_MBC_12(7)2119.pdf (1.409Mb)
    Issue Date
    2001-07-01
    Author
    Jung, M. Katherine
    Prigozhina, Natalie L.
    Oakley, C. Elizabeth
    Nogales, Eva
    Oakley, Berl R.
    Publisher
    American Society for Cell Biology
    Type
    Article
    Article Version
    Scholarly/refereed, publisher version
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    Abstract
    We have created 41 clustered charged-to-alanine scanning mutations of the mipA, γ-tubulin, gene of Aspergillus nidulans and have created strains carrying these mutations by two-step gene replacement and by a new procedure, heterokaryon gene replacement. Most mutant alleles confer a wild-type phenotype, but others are lethal or conditionally lethal. The conditionally lethal alleles exhibit a variety of phenotypes under restrictive conditions. Most have robust but highly abnormal mitotic spindles and some have abnormal cytoplasmic microtubule arrays. Two alleles appear to have reduced amounts of γ-tubulin at the spindle pole bodies and nucleation of spindle microtubule assembly may be partially inhibited. One allele inhibits germ tube formation. The cold sensitivity of two alleles is strongly suppressed by the antimicrotubule agents benomyl and nocodazole and a third allele is essentially dependent on these compounds for growth. Together our data indicate that γ-tubulin probably carries out functions essential to mitosis and organization of cytoplasmic microtubules in addition to its well-documented role in microtubule nucleation. We have also placed our mutations on a model of the structure of γ-tubulin and these data give a good initial indication of the functionally important regions of the molecule.
    Description
    This is the publisher's version, also available electronically from "www.molbiolcell.org".
    URI
    http://hdl.handle.net/1808/17462
    DOI
    https://doi.org/10.1091/mbc.12.7.2119
    ISSN
    1059-1524
    Collections
    • Molecular Biosciences Scholarly Works [526]
    Citation
    Jung, M., Prigozhina, N., Oakley, C., Nogales, E., & Oakley, B. (2001). Alanine-scanning mutagenesis of Aspergillus γ-tubulin yields diverse and novel phenotypes. Moleculary Biology of the Cell, 12(7), 2119-2136. http://www.dx.doi.org/10.1091/mbc.12.7.2119

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    Contact KU ScholarWorks
    785-864-8983
    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    785-864-8983

    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    Image Credits
     

     

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