NTNU NTNU, Norwegian University of Science and Technology www.ntnu.no/home The human 20-kDa 5'-(CGG)(n)-3'-binding protein is targeted to the nucleus and affects the activity of the FMR1 promoter. The human 20-kDa 5'-(CGG)(n)-3'-binding protein is targeted to the nucleus and affects the activity of the FMR1 promoter. J. Biol. Chem. (0021-9258) 2000 imex curation Müller-Hartmann H., Deissler H., Naumann F., Schmitz B., Schröer J., Doerfler W. 21-Jul-2014: Contacted by IntAct-Help. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Müller-Hartmann H., Deissler H., Naumann F., Schmitz B., Schröer J., Doerfler W. Accepted 2014-JUL-10 by ORCHARD J. Biol. Chem. (0021-9258) 2000 imex curation The human 20-kDa 5'-(CGG)(n)-3'-binding protein is targeted to the nucleus and affects the activity of the FMR1 promoter. The human 20-kDa 5'-(CGG)(n)-3'-binding protein is targeted to the nucleus and affects the activity of the FMR1 promoter. J. Biol. Chem. (0021-9258) 2000 imex curation Müller-Hartmann H., Deissler H., Naumann F., Schmitz B., Schröer J., Doerfler W. 21-Jul-2014: Contacted by IntAct-Help. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 emsa supershift electrophoretic mobility supershift assay emsa supershift weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Müller-Hartmann H., Deissler H., Naumann F., Schmitz B., Schröer J., Doerfler W. Accepted 2014-JUL-10 by ORCHARD J. Biol. Chem. (0021-9258) 2000 imex curation cgbp1_human CGG triplet repeat-binding protein 1 p20-CGGBP DNA-binding protein 20 kDa CGG-binding protein CGGBP1 CGGBP protein protein human Homo sapiens Human MERFVVTAPPARNRSKTALYVTPLDRVTEFGGELHEDGGKLFCTSCNVVLNHVRKSAISDHLKSKTHTKRKAEFEEQNVRKKQRPLTASLQCNSTAQTEKVSVIQDFVKMCLEANIPLEKADHPAVRAFLSRHVKNGGSIPKSDQLRRAYLPDGYENENQLLNSQDC 1AF69CDB885BB8AD rna28s_gene_probe Human ribosomal DNA complete repeating unit ds dna double stranded deoxyribonucleic acid ds DNA ds dna human Homo sapiens Human GCGCGGGTCGGGGCGGCGGCGGCGGCGGCGGTGGCGGCGGCGGCGGGGGCGGCGGGACCG fragment 823 from the human 28 S rRNA gene; nucleotides 8758–8817, GenBankTMaccession number U13369 fmr1_human_gene Fragile X mental retardation protein 1 FMR1 gene gene human Homo sapiens Human cggbp1-rna28s 1 4 unspecified role unspecified role bait bait radiolabel 32p radiolabel 32p radiolabel P32 32P 32P P32 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 3 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain human CGGBP1 wild type full length was used; species ascertained from PMID:9201980 which reports cloning of CGGBP1 from human-hela cells several CGGBP1 deletions mutants were also assayed in the same experimenta, all had strongly reduced binding physical association physical association Fig 1C Q9UFW8 inhibits expression at RNA level of ENSG00000102081 the DNA-probe used in the emsa interaction detection (rna28s) contains the same 5′-(CGG)n-3′ repeats as those found in the FMR1 region assayed in the reporter gene assay (see causality statement) CGGBP1 (Q9UFW8) decreases expression at mRNA level of FMR1 (ENSG00000102081) as ascertained by reporter gene assay in human-hela upon CGGBP1 overexpression rna28s-cggbp1 2 5 unspecified role unspecified role bait bait radiolabel 32p radiolabel 32p radiolabel P32 32P 32P P32 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 3 unspecified role unspecified role prey prey 'endogeneous' CGGBP1 in human-hela was assayed (native human-hela nuclear extracts) physical association physical association Fig 5D Q9UFW8 decreases expression at RNA level of ENSG00000102081 the DNA-probe used in the emsa interaction detection (rna28s) contains the same 5′-(CGG)n-3′ repeats as those found in the FMR1 region assayed in the reporter gene assay (see causality statement) CGGBP1 (Q9UFW8) decreases expression at mRNA level of FMR1 (ENSG00000102081) as ascertained by reporter gene assay in human-hela upon CGGBP1 overexpression