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Molecular and Cellular Biology, March 1999, p. 2389-2399, Vol. 19, No. 3
Department of Microbiology and Institute for
Cellular and Molecular Biology, The University of Texas at Austin,
Austin, Texas
Received 8 September 1998/Returned for modification 13 October
1998/Accepted 4 November 1998
A mutation in NMD3 was found to be lethal in the
absence of XRN1, which encodes the major cytoplasmic
exoribonuclease responsible for mRNA turnover. Molecular genetic
analysis of NMD3 revealed that it is an essential gene
required for stable 60S ribosomal subunits. Cells bearing a
temperature-sensitive allele of NMD3 had decreased levels
of 60S subunits at the nonpermissive temperature which resulted in the
formation of half-mer polysomes. Pulse-chase analysis of rRNA
biogenesis indicated that 25S rRNA was made and processed with kinetics
similar to wild-type kinetics. However, the mature RNA was rapidly
degraded, with a half-life of 4 min. Nmd3p fractionated as a
cytoplasmic protein and sedimented in the position of free 60S subunits
in sucrose gradients. These results suggest that Nmd3p is a cytoplasmic
factor required for a late cytoplasmic assembly step of the 60S subunit
but is not a ribosomal protein. Putative orthologs of Nmd3p exist in
Drosophila, in nematodes, and in archaebacteria but not in
eubacteria. The Nmd3 protein sequence does not contain readily
recognizable motifs of known function. However, these proteins all have
an amino-terminal domain containing four repeats of Cx2C,
reminiscent of zinc-binding proteins, implicated in nucleic acid
binding or protein oligomerization.
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
NMD3 Encodes an Essential Cytoplasmic
Protein Required for Stable 60S Ribosomal Subunits in
Saccharomyces cerevisiae
*
Corresponding author. Mailing address: Department of
Microbiology, The University of Texas at Austin, Austin, TX 78712-1095. Phone: (512) 475-6350. Fax: (512) 471-7088. E-mail:
arlen{at}mail.utexas.edu.
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