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Originally published In Press as doi:10.1074/jbc.M005947200 on October 19, 2000

J. Biol. Chem., Vol. 276, Issue 2, 915-923, January 12, 2001
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Molecular Cloning and Characterization of a Novel Regulator of G-protein Signaling from Mouse Hematopoietic Stem Cells*

In-Kyung ParkDagger , Christopher A. Klug§, Kaijun Li§, Libuse Jerabek§, Linheng Li||**, Masakatsu NanamoriDagger Dagger , Richard R. NeubigDagger Dagger , Leroy Hood||§§, Irving L. Weissman§, and Michael F. ClarkeDagger ¶¶

From the Dagger  Department of Internal Medicine, Division of Hematology and Oncology, University of Michigan, Ann Arbor, Michigan 48109, the § Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, the || Department of Molecular Biotechnology, University of Washington, Seattle, Washington 98195, and the Dagger Dagger  Department of Pharmacology, University of Michigan, Ann Arbor, Michigan 48109

A novel regulator of G-protein signaling (RGS) has been isolated from a highly purified population of mouse long-term hematopoietic stem cells, and designated RGS18. It has 234 amino acids consisting of a central RGS box and short divergent NH2 and COOH termini. The calculated molecular weight of RGS18 is 27,610 and the isoelectric point is 8.63. Mouse RGS18 is expressed from a single gene and shows tissue specific distribution. It is most highly expressed in bone marrow followed by fetal liver, spleen, and then lung. In bone marrow, RGS18 level is highest in long-term and short-term hematopoietic stem cells, and is decreased as they differentiate into more committed multiple progenitors. The human RGS18 ortholog has a tissue-specific expression pattern similar to that of mouse RGS18. Purified RGS18 interacts with the alpha  subunit of both Gi and Gq subfamilies. The results of in vitro GTPase single-turnover assays using Galpha i indicated that RGS18 accelerates the intrinsic GTPase activity of Galpha i. Transient overexpression of RGS18 attenuated inositol phosphates production via angiotensin receptor and transcriptional activation through cAMP-responsive element via M1 muscarinic receptor. This suggests RGS18 can act on Gq-mediated signaling pathways in vivo.

* This work was supported by Grant DK53074-04 from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF302685.

Present address: Dept. of Microbiology, Div. of Developmental and Clinical Immunology, University of Alabama at Birmingham, Birmingham, AL 35294.

** Present address: Stowers Institute for Medical Research, 1000 E. 50th St., Kansas City, MO 64110.

§§ Present address: Institute for Systems Biology, Seattle, WA 98105.

¶¶ To whom correspondence should be addressed: Dept. of Internal Medicine, Div. of Hematology and Oncology, University of Michigan, CCGC 4431, 1500 E. Medical Center Dr., Ann Arbor, MI 48109. Tel.: 734-647-3428; Fax: 734-647-9654; E-mail: mclarke@umich.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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