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Molecular and Cellular Biology, November 2000, p. 7943-7954, Vol. 20, No. 21
Institut de Génétique
Moléculaire, CNRS UMR 5535, 34000 Montpellier, France
Received 24 April 2000/Returned for modification 20 June
2000/Accepted 10 August 2000
In mammals, nuclear localization of U-snRNP particles requires the
snRNA hypermethylated cap structure and the Sm core complex. The nature
of the signal located within the Sm core proteins is still unknown,
both in humans and yeast. Close examination of the sequences of the
yeast SmB, SmD1, and SmD3 carboxyl-terminal domains reveals the
presence of basic regions that are reminiscent of nuclear localization
signals (NLSs). Fluorescence microscopy studies using green fluorescent
protein (GFP)-fusion proteins indicate that both yeast SmB and SmD1
basic amino acid stretches exhibit nuclear localization properties.
Accordingly, deletions or mutations in the NLS-like motifs of SmB and
SmD1 dramatically reduce nuclear fluorescence of the GFP-Sm mutant
fusion alleles. Phenotypic analyses indicate that the NLS-like motifs
of SmB and SmD1 are functionally redundant: each NLS-like motif can be
deleted without affecting yeast viability whereas a simultaneous
deletion of both NLS-like motifs is lethal. Taken together, these
findings suggest that, in the doughnut-like structure formed by the Sm core complex, the carboxyl-terminal extensions of Sm proteins may form
an evolutionarily conserved basic amino acid-rich protuberance that
functions as a nuclear localization determinant.
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Copyright © 2000, American Society for Microbiology. All rights reserved.
Functional Characterization of Nuclear
Localization Signals in Yeast Sm Proteins
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