In vivo protein-DNA interactions at human DNA replication origin.

• DOI: 10.1073/pnas.93.4.1498
• Publication Date: 2002-07-26T14:34:16Z
• Authors: D S Dimitrova; M Giacca; F Demarchi; G Biamonti; S Riva; A Falaschi
• Keywords: DNA footprinting; Footprinting; HMG-box

Protein-DNA interactions were studied in vivo at the region containing a human DNA replication origin, located 3' end of lamin B2 gene and partially overlapping promoter another gene, downstream. DNase I treatment nuclei isolated from both exponentially growing nonproliferating HL-60 cells showed that this has an altered, highly accessible, chromatin structure. High-resolution analysis protein-DNA 600-bp area encompassing origin was carried out by footprinting technique based on ligation-mediated polymerase chain reaction. In cells, footprints sequences homologous to binding sites for known transcription factors (members basic-helix-loop-helix family, nuclear respiratory factor 1, Sp1, upstream factor) detected corresponding downstream gene. Upon conversion nonproliferative state, reduction intensity these observed paralleled diminished transcriptional activity genomic area. addition protections, close correspondence initiation site, prominent footprint extended over 70 nucleotides one strand only. This absent indicating specific interaction might be involved process activation.