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PMID: 16251348 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The homologous putative GTPases Grn1p from fission yeast and the human GNL3L are required for growth and play a role in processing of nucleolar pre-rRNA.

Molecular biology of the cell ·Vol. 17 ·No. 1 ·2006-01-00 ·Pages 460-74

Du X, Rao MR, Chen XQ, Wu W, Mahalingam S, Balasundaram D

Abstract

Grn1p from fission yeast and GNL3L from human cells, two putative GTPases from the novel HSR1_MMR1 GTP-binding protein subfamily with circularly permuted G-motifs play a critical role in maintaining normal cell growth. Deletion of Grn1 resulted in a severe growth defect, a marked reduction in mature rRNA species with a concomitant accumulation of the 35S pre-rRNA transcript, and failure to export the ribosomal protein Rpl25a from the nucleolus. Deleting any of the Grn1p G-domain motifs resulted in a null phenotype and nuclear/nucleolar localization consistent with the lack of nucleolar export of preribosomes accompanied by a distortion of nucleolar structure. Heterologous expression of GNL3L in a Deltagrn1 mutant restored processing of 35S pre-rRNA, nuclear export of Rpl25a and cell growth to wild-type levels. Genetic complementation in yeast and siRNA knockdown in HeLa cells confirmed the homologous proteins Grn1p and GNL3L are required for growth. Failure of two similar HSR1_MMR1 putative nucleolar GTPases, Nucleostemin (NS), or the dose-dependent response of breast tumor autoantigen NGP-1, to rescue deltagrn1 implied the highly specific roles of Grn1p or GNL3L in nucleolar events. Our analysis uncovers an important role for Grn1p/GNL3L within this unique group of nucleolar GTPases.

MeSH Terms
Active Transport, Cell Nucleus Amino Acid Sequence Cell Nucleolus/genetics,metabolism Cell Proliferation Conserved Sequence GTP Phosphohydrolases/chemistry,classification,genetics,metabolism GTP-Binding Proteins Gene Deletion HeLa Cells Humans Models, Molecular Mutation/genetics Nuclear Proteins Protein Binding Protein Structure, Tertiary RNA Precursors/genetics Ribosomal Proteins/genetics,metabolism Schizosaccharomyces/cytology,enzymology,genetics Schizosaccharomyces pombe Proteins/chemistry,classification,genetics,metabolism Sequence Alignment
Chemicals
GNL3L protein, human Nuclear Proteins RNA Precursors Ribosomal Proteins Schizosaccharomyces pombe Proteins GTP Phosphohydrolases GTP-Binding Proteins Grn1 protein, S pombe
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Du Xianming
Laboratory of Nucleopore Biology, Institute of Molecular and Cell Biology, National University of Singapore, Singapore 117609, Singapore.
Rao Malireddi R K Subba
Chen Xue Qin
Wu Wei
Mahalingam Sundarasamy
Balasundaram David
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-09-0848
Published
2006-01-00
Epub
2005-00-26
Pages
460-74
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1345682
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e05-09-0848 PMC Full Text View journal details
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