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Inactivation of the yeast Sen1 protein affects the localization of nucleolar proteins

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Abstract

A mutation in the Saccharomyces cerevisiae SEN1 gene causes accumulation of end-matured, intron-containing pre-tRNAs. Cells containing the thermosensitive sen1-1 mutation exhibit reduced tRNA splicing endonuclease activity. However, Sen1p is not the catalytic subunit of this enzyme. We have used Sen1p-specific antibodies for cell fractionation studies and immunofluorescent microscopy and determined that Sentp is a low abundance protein of about 239 kDa. It localizes to the nucleus with a granular distribution. We verified that a region in SEN1 containing a putative nuclear localization signal sequence (NLS) is necessary for nuclear targeting. Furthermore, we found that inactivation of Sen1p by temperature shift of a strain carrying sen1-1 leads to mislocalization of two nucleolar proteins, Nopt and Ssb1 Possible mechanisms are discussed for several related nuclear functions of Sen1p, including tRNA splicing and the maintenance of a normal crescent-shaped nucleolus.

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Authors and Affiliations

  1. Laboratories of Genetics and Molecular Biology, University of Wisconsin, 1525 Linden Drive, 53706, Madison, WI, USA

    Doris Ursic & Michael R. Culbertson

  2. Department of Biology, University of North Carolina, 27599, Chapel Hill, NC, USA

    Douglas J. DeMarini

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  1. Doris Ursic
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  2. Douglas J. DeMarini
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  3. Michael R. Culbertson
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Communicated by C. P. Hollenberg

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Ursic, D., DeMarini, D.J. & Culbertson, M.R. Inactivation of the yeast Sen1 protein affects the localization of nucleolar proteins. Molec. Gen. Genet. 249, 571–584 (1995). https://doi.org/10.1007/BF00418026

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  • DOI: https://doi.org/10.1007/BF00418026

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