title: The primary function of a redundant Sp1 binding site in the mouse aprt gene promoter is to block epigenetic gene inactivation.
abstract: The promoter region of the mouse adenine phosphoribosyltransferase (aprt) gene contains one non-consensus Sp1 binding site at its 5' end followed by three consensus Sp1 binding sites. The two 3'-most binding sites are sufficient for maximal expression of aprt , suggesting that the non-consensus and consensus binding sites at the 5' end are redundant. However, the two 3' sites are not sufficient to block epigenetic inactivation, which led to the hypothesis that the redundant consensus and/or non-consensus 5' Sp1 binding sites are required to block inactivation events. To test this hypothesis, promoter region constructs were made in which the two 5' Sp1 binding sites were mutated alone or in tandem, and then each construct was tested for its ability to withstand epigenetic inactivation. A cis -acting methylation center that is normally located 1.2 kb upstream of the promoter was used to induce inactivation. The results demonstrate that the presence of the redundant consensus Sp1 binding site is required to block methylation-associated gene inactivation. Therefore, the Sp1 binding sites comprising the mouse aprt promoter have evolved two distinct functions, one to promote transcription and the other to block epigenetic inactivation.
authors.affiliation: Department of Pathology, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: P
authors.initials: P
authors.lastname: Mummaneni
authors.forename: P
authors.initials: P
authors.lastname: Yates
authors.forename: J
authors.initials: J
authors.lastname: Simpson
authors.forename: J
authors.initials: J
authors.lastname: Rose
authors.forename: M S
authors.initials: MS
authors.lastname: Turker
mesh_terms: D000225:Adenine; D000228:Adenine Phosphoribosyltransferase / Q000235:genetics*; D000818:Animals; D001377:Azaserine; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D014162:Transfection
chemical_list: D017931:DNA Primers; D016329:Sp1 Transcription Factor; D001377:Azaserine; D004247:DNA; D000228:Adenine Phosphoribosyltransferase; D000225:Adenine