Alternative splicing of CD45 pre-mRNA is uniquely obedient to conditions in lymphoid cells.
The leucocyte common antigen (LCA or CD45) consists of various isoforms generated by alternative splicing of variable exons 4, 5 and 6 (or A, B and C). To follow splicing behaviour in different cell types we developed a human CD45 mini-gene and analysed its expression in transfected cell lines and transgenic mouse tissues. In Cos-1, HeLa and 3T3 cells we found distinct expression patterns which could only be modulated slightly by protein synthesis inhibitors but not by variation in culture conditions like pH, serum concentration and cell density, or by stimulation with phorbol ester (TPA). In all non-lymphoid transgenic tissues the default splicing pattern (CD45R0) was found, while the expression profile in lymphoid cells, where all eight isoforms are present, mimics that of the endogenous mouse LCA gene products. Next, to examine the factors involved in alternative exon use we analysed the expression pattern of members of the family of SR proteins, well known splicing regulators with arginine/serine-rich (R/S) domains. Cell lines expressed variable levels of SRp75, SRp30 and SRp20 and constant amounts of SRp40. Mouse tissues expressed large amounts of SRp75, SRp55 and SRp40, additional expression of SRp30s and SRp20 was restricted to lymphoid tissues. Therefore, SRp30 and SRp20 may contribute to forming the appropriate cellular conditions for alternative use of CD45 exons 4-6 in the haematopoietic compartment.
ten Dam
G B
GB
Department of Cell Biology, Faculty of Medical Sciences, University of Nijmegen, The Netherlands. g.tendam@bioch.kun.nl
Wieringa
B
B
Poels
L G
LG
D017398:Alternative Splicing; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis / Q000235:genetics*; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D009687:Nuclear Proteins / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010957:Plasmids; D020033:Protein Isoforms / Q000235:genetics*; D012322:RNA Precursors / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000378:metabolism; D000068103:Serine-Arginine Splicing Factors; D014162:Transfection
D009687:Nuclear Proteins; D010750:Phosphoproteins; D020033:Protein Isoforms; D012322:RNA Precursors; D016601:RNA-Binding Proteins; C491388:SRSF3 protein, human; C081934:SRSF4 protein, human; C106161:Srsf3 protein, mouse; D000068103:Serine-Arginine Splicing Factors; D017493:Leukocyte Common Antigens