Cloning and functional analysis of the hematopoietic cell-specific phospholipase C(gamma)2 promoter.
Phospholipase C(gamma)2 (PLCgamma2) is a phospholipid-converting enzyme which, upon receptor stimulation, is activated within membrane-bound signalling complexes. In contrast to the highly ubiquitous PLCgamma1, PLCgamma2 is expressed predominantly in B-lymphocytes. Associated with antigen-coupling receptors it is activated by tyrosine phosphorylation after the triggering of B-cell surface immunoglobulin. We have cloned and sequenced the human PLCgamma2 promoter. Primer extension analysis reveals the existence of a major transcriptional start site. The TATA-less promoter contains G+C-rich stretches with a cluster of contiguous SP1 consensus sites, an NF1, and an AP2 site between bp -220 to -70. A construct containing the region from -189 to +78 confers full promoter activity, as shown by fusion to a luciferase reporter gene construct. The distal part of the promoter between bp -662 to -293 containing an SRE, EBF and CACCC box contributed negatively to promoter activity in the B-cell line Raji but not in three adherent cell lines. In Raji cells, PLCgamma2 mRNA is expressed at low levels with a half life greater than 4 h. After treatment with serum, TPA, retinoic acid, or with 5-azacytidine increased levels of PLCgamma2 mRNA were induced in B-cells.
Kang
J S
JS
Mount Sinai Medical School, Department of Biochemistry, New York, NY 10029, USA.
Kohlhuber
F
F
Hug
H
H
Marmé
D
D
Eick
D
D
Ueffing
M
M
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D001853:Bone Marrow / Q000378:metabolism*; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004789:Enzyme Activation; D006801:Humans; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D051966:Phospholipase C gamma; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D010738:Type C Phospholipases / Q000235:genetics* / Q000378:metabolism
D004274:DNA, Recombinant; D007527:Isoenzymes; D010738:Type C Phospholipases; D051966:Phospholipase C gamma