title: Characterization of the 5'-flanking region of the human DNA helicase B (HELB) gene and its response to trans-Resveratrol.
abstract: Human DNA helicase B (HELB/HDHB) regulates DNA replication through association with human DNA polymerase α-primase. In the present study, an 866-base pair (bp) of the 5'-flanking region of the human HELB gene-containing Luciferase (Luc) reporter plasmid, pHDHB-Luc was transfected into various cell lines and Luc activity was analyzed. Deletion analyses revealed that a 121-bp containing the major transcription start site (TSS) was essential for the basal promoter activity in all tested cells. TF-SEARCH analysis indicated that GC-box/Sp1 and duplicated GGAA-motifs containing putative STAT-x and c-ETS binding sites are located close to the TSS. Furthermore, chromatin immunoprecipitation (ChIP) analysis showed that PU.1 and Sp1 bind to the 121-bp region. Reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot analyses showed the HELB gene and protein expression was up-regulated by trans-Resveratrol (Rsv) treatment in HeLa S3 cells. Moreover, transfection experiment indicated that mutations on the GC-boxes and the duplicated GGAA-motif greatly reduced promoter activity and the response to Rsv in HeLa S3 cells. These results suggest that Rsv, which is a natural compound that has been found to elongate the lifespan of various organisms, regulates HELB promoter activity through co-operation of the GC-boxes and the duplicated GGAA-motif in the 121-bp.
authors.affiliation: Department of Gene Regulation, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda-shi, Chiba-ken 278-8510, Japan.|Research Center for RNA Science, RIST. Tokyo University of Science, Noda-shi, Chiba-ken 278-8510, Japan.
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Uchiumi
authors.affiliation: Department of Gene Regulation, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda-shi, Chiba-ken 278-8510, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Arakawa
authors.affiliation: Department of Gene Regulation, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda-shi, Chiba-ken 278-8510, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Iwakoshi
authors.affiliation: Department of Gene Regulation, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda-shi, Chiba-ken 278-8510, Japan.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Ishibashi
authors.affiliation: Research Center for RNA Science, RIST. Tokyo University of Science, Noda-shi, Chiba-ken 278-8510, Japan.|Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda-shi, Chiba-ken 278-8510, Japan.
authors.forename: Sei-ichi
authors.initials: S
authors.lastname: Tanuma
mesh_terms: D024506:5' Flanking Region; D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004265:DNA Helicases / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D011401:Promoter Regions, Genetic; D059748:Proteolysis; D020218:Response Elements; D000077185:Resveratrol; D016329:Sp1 Transcription Factor / Q000378:metabolism; D013267:Stilbenes / Q000494:pharmacology*; D024363:Transcription Initiation Site
chemical_list: D016329:Sp1 Transcription Factor; D013267:Stilbenes; C484263:HELB protein, human; D004265:DNA Helicases; D000077185:Resveratrol