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PMID: 9271378 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Saccharomyces cerevisiae Nip7p is required for efficient 60S ribosome subunit biogenesis.

Molecular and cellular biology ·Vol. 17 ·No. 9 ·1997-09-00 ·Pages 5001-15

Zanchin NI, Roberts P, DeSilva A, Sherman F, Goldfarb DS

Abstract

The Saccharomyces cerevisiae temperature-sensitive (ts) allele nip7-1 exhibits phenotypes associated with defects in the translation apparatus, including hypersensitivity to paromomycin and accumulation of halfmer polysomes. The cloned NIP7+ gene complemented the nip7-1 ts growth defect, the paromomycin hypersensitivity, and the halfmer defect. NIP7 encodes a 181-amino-acid protein (21 kDa) with homology to predicted products of open reading frames from humans, Caenorhabditis elegans, and Arabidopsis thaliana, indicating that Nip7p function is evolutionarily conserved. Gene disruption analysis demonstrated that NIP7 is essential for growth. A fraction of Nip7p cosedimented through sucrose gradients with free 60S ribosomal subunits but not with 80S monosomes or polysomal ribosomes, indicating that it is not a ribosomal protein. Nip7p was found evenly distributed throughout the cytoplasm and nucleus by indirect immunofluorescence; however, in vivo localization of a Nip7p-green fluorescent protein fusion protein revealed that a significant amount of Nip7p is present inside the nucleus, most probably in the nucleolus. Depletion of Nip7-1p resulted in a decrease in protein synthesis rates, accumulation of halfmers, reduced levels of 60S subunits, and, ultimately, cessation of growth. Nip7-1p-depleted cells showed defective pre-rRNA processing, including accumulation of the 35S rRNA precursor, presence of a 23S aberrant precursor, decreased 20S pre-rRNA levels, and accumulation of 27S pre-rRNA. Delayed processing of 27S pre-rRNA appeared to be the cause of reduced synthesis of 25S rRNA relative to 18S rRNA, which may be responsible for the deficit of 60S subunits in these cells.

MeSH Terms
Alleles Amino Acid Sequence Cloning, Molecular Fungal Proteins/biosynthesis,genetics,physiology Gene Deletion Genes, Fungal Molecular Sequence Data Mutagenesis RNA Precursors/metabolism Restriction Mapping Ribosomal Proteins/genetics,metabolism Ribosomes/metabolism,ultrastructure Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins
Chemicals
Fungal Proteins NIP7 protein, S cerevisiae RNA Precursors Ribosomal Proteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Zanchin N I
Department of Biology, University of Rochester, New York 14627, USA.
Roberts P
DeSilva A
Sherman F
Goldfarb D S
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.9.5001
Published
1997-09-00
Pages
5001-15
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232351
Subset
IM
Grants
NIGMS NIH HHS · R01GM12702 · United States
External Links
PubMed source DOI: 10.1128/MCB.17.9.5001 PMC Full Text View journal details
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