The snRNP-associated U1A levels change following IL-6 stimulation of human B-cells.
The U1A protein can be found both in a small-ribonucleoprotein particle (snRNP) that contains U1 RNA, or in a distinctive fraction, free of the snRNP, the SF-A complex. Both components have been shown to influence post- or co-transcriptional RNA processing reactions in HeLa cells. Since U1A may influence the processing of the immunoglobulin heavy chain pre-mRNA in B-cells, we wanted to see if the levels of U1A in either of its two forms changed following IL-6 stimulation to IgM secretion. Using antibodies that specifically recognize the two forms of U1A, snRNP-associated and snRNP-free, we found that approximately 16% of U1A is in the SF-A form in B-cells. We measured the levels of U1A protein in its two states in human B-cell lines both by flow cytometry and exhaustive immunoprecipitations. We found a significant decrease in the amount of snRNP-associated U1A following cytokine stimulation that correlates with the change-over to the secretory-specific poly(A) site use in the SKW 6.4 cell line. Meanwhile, the number of U1A molecules in the SF-A fraction of the pool remains nearly constant following induction to secretion. Our results suggest that the changing level of U1A in the snRNP fraction may be important for influencing Ig heavy chain mRNA processing.
Milcarek
Christine
C
Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA. milcarek@pitt.edu
Martincic
Kathleen
K
Chung-Ganster
Ling-Hsiu
LH
Lutz
Carol S
CS
D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D015850:Interleukin-6 / Q000494:pharmacology; D012323:RNA Processing, Post-Transcriptional; D016601:RNA-Binding Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000235:genetics / Q000378:metabolism*
D007143:Immunoglobulin Heavy Chains; D015850:Interleukin-6; D016601:RNA-Binding Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear; C075339:U1A protein