Previous studies have indicated that transcription of germ-line (GL) C-H genes is necessary to obtain immunoglobulin (Ig) class switching. We report here a correlation between proliferation, switching and GL transcripts. S mu-S gamma 1 switch recombination in lipopolysaccharide (LPS) + interleukin-4 (IL-4)-activated mouse B cells was assayed by a digestion-circularization polymerase chain reaction. Switching to gamma 1 is reduced upon inhibition of DNA synthesis with hydroxyurea (HU) or aphidicholin (AC). Incubation of activated B cells with HU severely reduces steady-state levels of GL gamma 1 and epsilon RNA. By utilizing elutriation to synchronize B cell blasts in different phases of the cell cycle, it was found that GL gamma 1 transcripts are mainly expressed in G(1) and S phases, but not in G(0). Using the electrophoretic mobility shift assay, we characterized two major LPS-induced complexes, which bind to the GL gamma 1 promoter and are expressed at levels which correlate with the amount of LPS-induced DNA synthesis. Furthermore, the intensity of the complexes is reduced when cells are arrested with the DNA synthesis inhibitors HU or AC. Elutriation experiments revealed that the complexes are expressed in G(1) and S, but not in G(0). They bind to an Ets consensus element near the major initiation sites used in proliferating cells. The possible implications of these findings for Ig isotype switching are discussed.
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UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235
BERTON, MT
UHR, JW
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UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235
UHR, JW
VITETTA, ES
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UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235
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UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235
BERTON, MT
UHR, JW
论文数: 0引用数: 0
h-index: 0
机构:
UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235
UHR, JW
VITETTA, ES
论文数: 0引用数: 0
h-index: 0
机构:
UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235