Chromatin remodeling occurs independent of transcription factor binding during 5-azacytidine reactivation of the human HPRT gene
- Univ. of Florida College of Medicine, Gainesville, FL (United States); and others
A novel system of differential gene expression in mammals is established during normal female embryogenesis by X chromosome inactivation. Studies of 5-aza-2{prime}-deoxycytidine (5aCdr)-induced reactivation of genes on the inactive human X chromosome strongly implicate DNA methylation in maintaining the transcriptional repression of discrete loci on the inactive X. During the process of 5aCdr-induced reactivation of the human hypoxanthine phosphoribosyltransferase (HPRT) gene on the inactive X chromosome, changes in nuclease sensitivity of chromatin in the 5{prime} region of the HPRT gene and HPRT mRNA levels have been analyzed from 0-72 hrs. after 5aCdr exposure. Increased nuclease sensitivity is first detectable at 6 hrs. and reaches a maximum at 24 hrs. after initial exposure to 5aCdr, while the appearance of HPRT mRNA levels is first detectable by RT-PCR at 24 hrs. and reaches a maximum of 48 hrs. after 5aCdr exposure. Thus, the change in chromatin structure of the 5{prime} region as a result of 5aCdr treatment appears to occur prior to active transcription of the gene. However, it is unclear if the remodeling of chromatin requires the binding of transcription factors to the 5{prime} region, or if the binding of transcription factors is only required for transcription of the HPRT gene. We now have assayed the binding of transcription factors to the 5{prime} region of the HPRT gene on the inactive X chromosome during 5aCdr reactivation. We find that the change in chromatin structure as a result of 5aCdr treatment occurs independent of transcription factor binding, and that the binding of factors is correlated with active transcription of the gene rather than remodeling of chromatin structure. These data suggest that the differential binding of transcriptional activators (and differential expression of the HPRT gene) to the active and inactive HPRT genes is modulated by the accessibility of their binding sites due to chromatin structure.
- OSTI ID:
- 133840
- Report Number(s):
- CONF-941009-; ISSN 0002-9297; TRN: 95:005313-0573
- Journal Information:
- American Journal of Human Genetics, Vol. 55, Issue Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
- Country of Publication:
- United States
- Language:
- English
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