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PMID: 9305759 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

SA-1, a nuclear protein encoded by one member of a novel gene family: molecular cloning and detection in hemopoietic organs.

Gene ·Vol. 195 ·No. 2 ·1997-08-22 ·Pages 151-9

Carramolino L, Lee BC, Zaballos A, Peled A, Barthelemy I, Shav-Tal Y, Prieto I, Carmi P, Gothelf Y, González de Buitrago G, Aracil M, Márquez G, Barbero JL, Zipori D

Abstract

We report the molecular cloning of a novel gene family. The first member of this family was cloned from a mouse lambda gt11 cDNA library using the B92 monoclonal antibody (mAb) raised against stromal cell extracts. This was followed by RACE-PCR using mRNA from the stromal cell line. A 4 kb cDNA was obtained encoding a unique protein sequence of 1258 aa, that we designate stromal antigen (SA)-1. The human SA-1 gene was cloned by homology from a thymus cDNA library and the sequence of the predicted protein was found to be highly homologous to the murine SA-1 (>98.9%). Another cDNA was cloned and the deduced protein (SA-2) was 71% homologous to SA-1. Northern blot and PCR analysis indicated that on the mRNA level the SA-1 gene is expressed in all tissues analyzed and probably encodes a single transcript. The identification of SA-1 protein in tissues and cells required combined immunoprecipitation and Western blotting using a polyclonal antiserum raised against a predicted peptide of SA-1 and the B92 mAb. Using this assay we identified a protein of about 120 kDa in hemopoietic organs. Subcellular fractionation indicated that SA-1 is a nuclear protein. Thus, despite the ubiquitous expression on the mRNA level, the protein was predominantly detected in hemopoietic organs and may therefore be controlled on a post-transcriptional level. The SA-1 gene described in this study is highly conserved between mouse and man. This implies a crucial function for this protein.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Blotting, Western Bone Marrow Cells/cytology,metabolism Cells, Cultured Cloning, Molecular Female Gene Expression Gene Library Humans Lymphocytes/metabolism Male Mice Mice, Inbred BALB C Molecular Sequence Data Neoplasms, Experimental/genetics,metabolism Nuclear Proteins/chemical synthesis,genetics,metabolism Peptides/chemical synthesis,immunology Polymerase Chain Reaction Precipitin Tests RNA, Messenger/genetics,metabolism Recombination, Genetic Sequence Alignment Sequence Analysis, DNA Sequence Homology, Amino Acid Stromal Cells Thymus Gland/metabolism Transcription, Genetic
Chemicals
Nuclear Proteins Peptides RNA, Messenger STAG1 protein, human
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Carramolino L
Pharmacia and UpJohn, R&D, Madrid, Spain.
Lee B C
Zaballos A
Peled A
Barthelemy I
Shav-Tal Y
Prieto I
Carmi P
Gothelf Y
González de Buitrago G
Aracil M
Márquez G
Barbero J L
Zipori D
Article Info
Journal
Gene
Abbr.
Gene
ISSN
0378-1119
Published
1997-08-22
Pages
151-9
Language
English
Region
Netherlands
NLM ID
7706761
Subset
IM
Databases
GENBANK
Z75330, Z75331, Z75332
Corrections
ErratumIn
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