Molecular Cloning, Sequence Analysis, Expression, and Tissue Distribution of Suppressin, a Novel Suppressor of Cell Cycle Entry

doi:10.1074/jbc.273.1.361

Molecular Cloning, Sequence Analysis, Expression, and Tissue Distribution of Suppressin, a Novel Suppressor of Cell Cycle Entry*

From the Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, Alabama 35294

Abstract

Suppressin (SPN) is an inhibitor of cell proliferation that was originally identified and purified to homogeneity from bovine pituitaries (LeBoeuf, R. D., Burns, J. N., Bost, K. L., and Blalock, J. E. (1990)J. Biol. Chem. 265, 158–165). In this report we have cloned the full-length cDNA encoding rat SPN and have identified the tissue distribution of SPN expression. The cDNA of SPN is 1882 nucleotides with a 1488-base coding region and 55 and 339 nucleotides of 5′- and 3′-untranslated sequences, respectively. Northern gel analysis of rat pituitary mRNA showed a single hybridizing species at ∼2 kilobases. Sequence analyses showed that the nucleotide and deduced amino acid sequences of SPN are novel and unrelated to any known vertebrate inhibitors of proliferation. However, the deduced amino acid sequence of SPN contains two domains that have extensive sequence identity with a recently cloned transcription activator inDrosophila, deformed epidermal autoregulatory factor-1 (DEAF-1, see Gross, C. T., and McGinnis, W. (1996) EMBO J.15, 1961–1970) suggesting that SPN represents a vertebrate cognate of deformed epidermal autoregulatory factor-1. Reverse transcriptase-polymerase chain reaction and immunohistochemical analyses showed that the SPN mRNA and the SPN protein are expressed in every tissue examined including testis, spleen, skeletal muscle, liver, kidney, heart, and brain suggesting that SPN may be involved in the control of proliferation in a variety of cell types.

Footnotes

↵* This research was supported in part by NCI Grant CA 54290 from the National Institutes of Health (to R. D. L.). Sequence analyses were performed through the computer facilities of the UAB-Center for AIDS Research supported in part by Grant P30 AI27767 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 GenBank™/EMBL Data Bank with accession number(s) .
↵‡ To whom correspondence should be addressed: Dept. of Physiology and Biophysics, BHSB 850, University of Alabama at Birmingham, Birmingham, AL 35294. Tel.: 205-934-4271; Fax: 205-934--6674.
↵§ Supported by a fellowship from the IPSEN Foundation of France. Present address: Unite INSERM 167, Institut Pasteur de Lille, 1, rue du Professeur Calmette, 59019 Lille Cedex, France.
↵¶ Present address: Dept. of Microbiology, University of Florida, Gainesville, FL 35294.
↵‖ Present address: Dept. of Chemistry, McNeese State University, Lake Charles, LA 70606.
↵1 The abbreviations used are: SPN, suppressin; RT, reverse transcription; PCR, polymerase chain reaction; PBS, phosphate-buffered saline; mAb, monoclonal antibody; Ab, antibody; RACE, 5′-rapid amplification of cDNA ends; bp, base pair(s); PAGE, polyacrylamide gel electrophoresis; ORF, open reading frame; FGF-1, fibroblast growth factor-1; EST, expression sequence-tagged; DEAF-1, deformed epidermal autoregulatory factor-1; PBMC, peripheral blood mononuclear cells; PNGase F, peptide-N ⁴-(N-acetyl-β-glucosaminyl)-asparagine amidase.
↵2 R. D. LeBoeuf, M. M. Green, J. E. Blalock, and J. D. Tauber, unpublished observations.
- Received January 22, 1997.
- Revision received July 27, 1997.
The American Society for Biochemistry and Molecular Biology, Inc.