TFIIIA binds with equal affinity to somatic and major oocyte 5S RNA genes.
Current models for the differential control of expression of Xenopus somatic and oocyte 5S RNA genes suggest that an impaired ability to bind TFIIIA contributes to the inactivation of oocyte 5S RNA genes in somatic cells. The somatic 5S RNA gene is transcribed more efficiently than the major oocyte 5S RNA gene in S-150 extracts of mature oocytes. However, this differential transcription efficiency is not determined simply by the relative affinity for binding of a positive transcription factor, TFIIIA. We have compared the abilities of somatic, major oocyte, and minor oocyte 5S RNA genes to bind TFIIIA using both a standard footprint competition assay and an indirect DNase protection assay. This indirect DNase protection assay permits the direct comparison of TFIIIA binding to two templates in one reaction. Both assay methods indicate that the major oocyte 5S RNA gene and the somatic 5S RNA gene bind TFIIIA with equal affinity. As a further control, we have confirmed earlier work indicating that the minor oocyte gene binds TFIIIA with a reduced affinity. Binding of TFIIIA to these three 5S RNA genes results in a different pattern of protection of each gene. We suggest that slight differences in the contacts between TFIIIA and the 5' border of the control region influence the ability of additional transcription factors to bind to the TFIIIA:5S DNA complex.
McConkey
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GA
Department of Pharmacological Sciences, State University of New York, Stony Brook 11794.
Bogenhagen
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DF
D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000235:genetics / Q000378:metabolism; D003851:Deoxyribonucleases; D005260:Female; D005796:Genes; D066298:In Vitro Techniques; D009865:Oocytes / Q000378:metabolism; D012335:RNA, Ribosomal / Q000235:genetics*; D012341:RNA, Ribosomal, 5S / Q000235:genetics* / Q000378:metabolism; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014981:Xenopus
D012335:RNA, Ribosomal; D012341:RNA, Ribosomal, 5S; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors; D004247:DNA; D003851:Deoxyribonucleases