Molecular events during B lymphocyte differentiation. Induction of endogenous mouse mammary tumor proviral envelope transcripts after B cell stimulation.
We have identified a gene whose expression appears to be associated with a late stage in the differentiation of B lymphocytes into antibody secreting cells, as shown by using the inducible B cell lymphoma, CH12. Restriction mapping and partial sequencing of a cDNA clone isolated by subtraction analysis demonstrated that the clone, SC34, represents an envelope (env) gene transcript of a mouse mammary tumor virus (MMTV). In CH12 cells and in normal B cells, levels of MMTV RNA were increased after stimulation with LPS. The env transcript was the predominant MMTV RNA species and increased more dramatically than did levels of the genomic transcript. In differentiating CH12 cells, env transcripts increased as much as 20-fold above levels found in replicating, antibody nonsecreting CH12 cells. The major increase in expression appeared to be associated with B cell differentiation and not replication. By Southern blot analysis, only bands characteristic of endogenous proviruses were found in CH12, indicating that viral sequences were not amplified in this cell line. Restriction mapping indicated that the SC34 cDNA clone was a product of the Mtv-9 locus. Mtv-9 previously was shown to encode a complete MMTV provirus on chromosome 12, on which Ig heavy chain genes also are located. Increases in MMTV transcripts followed distinct kinetics and were quantitatively different from changes in immunoglobulin gene products. The expression of env RNA appears to more accurately reflect differentiation to antibody secretion in CH12 cells than does the expression of immunoglobulin gene transcripts.
Sharma
S
S
Department of Microbiology and Immunology, Duke University Medical Center, Durham, NC 27710.
King
L B
LB
Corley
R B
RB
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D005814:Genes, Viral; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D008213:Lymphocyte Activation; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011533:Proviruses / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis; D014158:Transcription, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000302:isolation & purification
D008070:Lipopolysaccharides; D012367:RNA, Viral; D014759:Viral Envelope Proteins