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Vol. 9, Issue 11, 3041-3055, November 1998
Department of Biochemistry and Molecular Biology, Pennsylvania
State University College of Medicine, Hershey, Pennsylvania 17033
To understand the factors specifically affecting tRNA nuclear
export, we adapted in situ hybridization procedures to locate endogenous levels of individual tRNA families in wild-type and mutant
yeast cells. Our studies of tRNAs encoded by genes lacking introns show
that nucleoporin Nup116p affects both poly(A) RNA and tRNA export,
whereas Nup159p affects only poly(A) RNA export. Los1p is similar to
exportin-t, which facilitates vertebrate tRNA export. A
los1 deletion mutation affects tRNA but not poly(A) RNA
export. The data support the notion that Los1p and exportin-t are
functional homologues. Because LOS1 is nonessential,
tRNA export in vertebrate and yeast cells likely involves factors in addition to exportin-t. Mutation of RNA1, which encodes
RanGAP, causes nuclear accumulation of tRNAs and poly(A) RNA.
Many yeast mutants, including those with the rna1-1
mutation, affect both pre-tRNA splicing and RNA export. Our studies of
the location of intron-containing pre-tRNAs in the
rna1-1 mutant rule out the possibility that this results
from tRNA export occurring before splicing. Our results also argue
against inappropriate subnuclear compartmentalization causing defects
in pre-tRNA splicing. Rather, the data support "feedback" of
nucleus/cytosol exchange to the pre-tRNA splicing machinery.
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