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PMID: 7799964 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Molecular cloning of a novel mitogen-inducible nuclear protein with a Ran GTPase-activating domain that affects cell cycle progression.

Molecular and cellular biology ·Vol. 15 ·No. 1 ·1995-01-00 ·Pages 552-60

Hattori M, Tsukamoto N, Nur-e-Kamal MS, Rubinfeld B, Iwai K, Kubota H, Maruta H, Minato N

Abstract

We have cloned a novel cDNA (Spa-1) which is little expressed in the quiescent state but induced in the interleukin 2-stimulated cycling state of an interleukin 2-responsive murine lymphoid cell line by differential hybridization. Spa-1 mRNA (3.5 kb) was induced in normal lymphocytes following various types of mitogenic stimulation. In normal organs it is preferentially expressed in both fetal and adult lymphohematopoietic tissues. A Spa-1-encoded protein of 68 kDa is localized mostly in the nucleus. Its N-terminal domain is highly homologous to a human Rap1 GTPase-activating protein (GAP), and a fusion protein of this domain (SpanN) indeed exhibited GAP activity for Rap1/Rsr1 but not for Ras or Rho in vitro. Unlike the human Rap1 GAP, however, SpanN also exhibited GAP activity for Ran, so far the only known Ras-related GTPase in the nucleus. In the presence of serum, stable Spa-1 cDNA transfectants of NIH 3T3 cells (NIH/Spa-1) hardly overexpressed Spa-1 (p68), and they grew as normally as did the parental cells. When NIH/Spa-1 cells were serum starved to be arrested in the G1/G0 phase of the cell cycle, however, they, unlike the control cells, exhibited progressive Spa-1 p68 accumulation, and following the addition of serum they showed cell death resembling mitotic catastrophes of the S phase during cell cycle progression. The results indicate that the novel nuclear protein Spa-1, with a potentially active Ran GAP domain, severely hampers the mitogen-induced cell cycle progression when abnormally and/or prematurely expressed. Functions of the Spa-1 protein and its regulation are discussed in the context of its possible interaction with the Ran/RCC-1 system, which is involved in the coordinated nuclear functions, including cell division.

Related Genes
Rap1GAP Spa-1
MeSH Terms
3T3 Cells Amino Acid Sequence Animals Base Sequence Cell Cycle Cloning, Molecular DNA Primers/chemistry Enzyme Activation GTP-Binding Proteins/metabolism GTPase-Activating Proteins Gene Expression Lymphocytes/cytology,metabolism Mice Mitogens/pharmacology Molecular Sequence Data Nuclear Proteins/genetics,metabolism,physiology RNA, Messenger/genetics Sequence Alignment Sequence Homology, Amino Acid ran GTP-Binding Protein rap GTP-Binding Proteins
Chemicals
DNA Primers GTPase-Activating Proteins Mitogens Nuclear Proteins RNA, Messenger SIPA1 protein, human Sipa1 protein, mouse GTP-Binding Proteins ran GTP-Binding Protein rap GTP-Binding Proteins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Hattori M
Department of Immunology and Cell Biology, Faculty of Medicine, Kyoto University, Japan.
Tsukamoto N
Nur-e-Kamal M S
Rubinfeld B
Iwai K
Kubota H
Maruta H
Minato N
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.1.552
Published
1995-01-00
Pages
552-60
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232010
Subset
IM
Databases
GENBANK
D11374
External Links
PubMed source DOI: 10.1128/MCB.15.1.552 PMC Full Text View journal details
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