Requirement of an ETS-binding element for transcription of the human lck type I promoter.
The requirement for cis-acting DNA sequences for transcriptional activity of the human lck type I promoter was investigated in two human cell lines that express type I transcripts, the leukemic T-cell line, Jurkat, and the colon carcinoma line, SW620. Transient transfection assays in Jurkat and SW620 cells revealed negative and positive cis-acting regulatory elements in the lck type I promoter between -570 and -480 and between -128 and -63 respectively. For the latter, a triple point mutation of a sequence, GCAGGAAGT, from -99 and -91 resulted in complete loss of lck type I promoter activity in both Jurkat and SW620 cells. In vitro binding assays indicated that this sequence, denoted the ETS-binding element or EBE, can interact with the lymphoid-specific transcription factor ETS-1. Thus, a protein(s) in the ETS family appears to be required for transcription of the lck type I promoter in T cells and may be important for the activation of the lck gene in human colon carcinoma.
Leung
S
S
Division of Cellular and Molecular Biology, Ontario Cancer Institute/Princess Margaret Hospital, Toronto, Canada.
McCracken
S
S
Ghysdael
J
J
Miyamoto
N G
NG
D000328:Adult; D001483:Base Sequence; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D066298:In Vitro Techniques; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; C494041:ETS1 protein, human; D009838:Oligodeoxyribonucleotides; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014157:Transcription Factors; D011505:Protein-Tyrosine Kinases; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)