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Published October 1987 | Published
Journal Article Open

The subnuclear localization of tRNA ligase in yeast

Abstract

Yeast tRNA ligase is an enzyme required for tRNA splicing. A study by indirect immune fluorescence shows that this enzyme is localized in the cell nucleus. At higher resolution, studies using indirect immune electron microscopy show this nuclear location to be primarily at the inner membrane of the nuclear envelope, most likely at the nuclear pore. There is a more diffuse, secondary location of ligase in a region of the nucleoplasm within 300 nm of the nuclear envelope. When the amount of ligase in the cell is increased, nuclear staining increases but staining of the nuclear envelope remains constant. This experiment indicates that there are a limited number of ligase sites at the nuclear envelope. Since the other tRNA splicing component, the endonuclease, has the characteristics of an integral membrane protein, we hypothesize that it constitutes the site for the interaction of ligase with the nuclear envelope.

Additional Information

© 1987 by The Rockefeller University Press. Received for publication 20 March 1987, and in revised form 4 June 1987. We thank Dr. Eric M. Phizicky for the yeast and E. coli strains used in this study. We also thank Drs. Eric Phizicky, Chris Greer, Brenda Fung, and Robert Singer for useful discussions and Dr. Jean Paul Revel for use of his Phillips 201B electron microscope. This work was supported by National Institutes of Health postdoctoral fellowship No. 5F32 GM1008-02 and American Cancer Society Grant No. NP302D (to J. Abelson).

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