Beta globin gene transcripts originating in the promoter region during early hexamethylene bisacetamide and dimethylsulfoxide induction of Friend erythroleukemia cells.
The beta-globin transcripts which are induced by dimethylsulfoxide (DMSO) and hexamethylene bisacetamide (HMBA) have been characterized in order to assess potential differences in their mechanisms of induction. Transcripts which initiate in the 5' flanking promoter region are likely indicators of promoter accessibility and were therefore characterized during the time course of induction with each inducer in Friend Erythroleukemia cells. S1 analysis with probes labeled at - 12 or +82 relative to the (+1) cap site showed no major differences between 5' ends of the upstream initiated transcripts in cells induced by DMSO or HMBA. We detected several upstream bands with each inducer corresponding to beta-globin transcripts with 5' ends between - 190 and -55 relative to the cap site and found that cells induced with DMSO and HMBA show a similar transcription response as measured by initiation in the 5' flanking region of the beta-globin gene. Interestingly, the upstream initiated transcripts reach their peak concentration levels much earlier in the time course of induction than do the mRNA transcripts with 5' ends at the major (+1) cap site. Northern blot analysis detected the upstream initiated transcripts as early as 16 hours after induction with DMSO, primarily in unprocessed large transcripts. We find that the promoter region containing transcripts constitute a higher percent of total beta-globin transcripts at the start of the induction and may therefore have an early function in the multistep induction process.
Weidner
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Department of Biochemistry University of Nevada Reno 89557.
Winicov
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D000081:Acetamides / Q000494:pharmacology*; D002460:Cell Line; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005622:Friend murine leukemia virus; D005786:Gene Expression Regulation; D005914:Globins / Q000235:genetics*; D004915:Leukemia, Erythroblastic, Acute; D007942:Leukemia, Experimental / Q000235:genetics; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D014158:Transcription, Genetic
D000081:Acetamides; D012333:RNA, Messenger; D005914:Globins; C014026:hexamethylene bisacetamide; D004121:Dimethyl Sulfoxide