The binding of a subunit of the general polyadenylation factor cleavage-polyadenylation specificity factor (CPSF) to polyadenylation sites changes during B cell development.
During the development of mouse B cells there is a regulated shift from the production of membrane (mb) to secretory-specific (sec) forms of immunoglobulin (Ig) mRNA. The mRNAs are produced from one gene that is alternatively processed at the 3' end. We have previously shown that there is an increase in polyadenylation efficiency accompanying the developmentally regulated shift to secretory-specific forms of Ig mRNA by DNA transfection experiments (1). When we look in vitro at nuclear extracts prepared from early/memory versus late stage/plasma B cells, we see cell stage-specific differences in the proteins which are crosslinked to poly(A) site-containing RNAs. Here we show that one of these proteins is the mouse homologue of 100 kDa subunit of Hela CPSF by immunoprecipitation and Western analysis of UV crosslinked material. The amount of 100 kDa protein and its mobility on two-dimensional gels do not change between the B cell stages. However, the binding of the 100 kDa polypeptide to poly(A) sites increases in the late stage/plasma cell lines relative to the binding seen in early/memory cell lines. The increased binding may reflect an increase in polyadenylation efficiency at the sec poly(A) site in plasma cells versus early/memory cells seen in vivo.
Edwalds-Gilbert
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Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, PA 15261-2072, USA.
Milcarek
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C
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D001665:Binding Sites; D002454:Cell Differentiation; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007156:Immunologic Memory; D066298:In Vitro Techniques; D051379:Mice; D010950:Plasma Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D011487:Protein Conformation; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D039221:mRNA Cleavage and Polyadenylation Factors
D007143:Immunoglobulin Heavy Chains; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D039221:mRNA Cleavage and Polyadenylation Factors