The insulin-like growth factor binding protein-1 (IGFBP-1) gene is highly expressed in fetal, perinatal, and regenerating liver. Up-regulation is transcriptionally mediated in regenerating liver and occurs in the first few minutes to hours after partial hepatectomy. In transgenic mice a 970-bp region from ؊776 to ؉151 of the IGFBP-1 promoter was sufficient for tissue-specific and induced expression of the gene in fetal and hepatectomized livers. However weak and/or poorly regulated expression in some transgenic lines suggested the existence of other regulatory regions. Here, genomic clones containing large regions 5Ј of the mouse IGFBP-1 gene sequence were isolated, subcloned, and sequenced. Deoxyribonuclease I (DNaseI) hypersensitivity analyses identified clusters of tissue-specific nucleasesensitive sites in the promoter region, ؊100 to ؊300, ؊2,300, ؊3,100, and ؊5,000 along with other weak sites. After partial hepatectomy, enhanced sensitivity and/or novel sites were detected in the ؊100/؊300, ؊5,000, and ؊3,100 regions, the promoter region remaining the most hypersensitive. A subset of these sites was present in fetal and perinatal livers. Novel tissue-specific sites that interacted with C/EBP and hepatic nuclear factor 3 (HNF3) transcription factors were identified in the ؊3,100 region. A hepatectomy-induced DNA binding complex containing the transcription factor USF1 was identified within the ؊100 to ؊300 region of the promoter. These results suggested that a complex array of tissue-specific and hepatic proliferationinduced transcription factors combine to regulate both the proximal promoter and more distal regulatory elements of the