PMID: 8958781Nov 1, 1996Paper

Transcription initiation of the rat insulin-like growth factor-I gene in hepatocyte primary culture

The Journal of Endocrinology
A Y KrishnaL S Phillips

Abstract

Transcription initiation in the insulin-like growth factor-I (IGF-I) gene is complex, involving multiple sites in two exons. While most transcripts are initiated in exon 1 in vivo, critical regulatory mechanisms are difficult to assess in intact animals. To examine the impact of insulin and growth hormone (GH) under more controlled conditions, we have studied the utilization of different exon 1 and exon 2 transcription-initiation sites in normal rat hepatocytes in primary culture. Normal rat hepatocytes were cultured for 48 h in serum-free medium, with insulin at 10(-6) or 10(-11) M, and with or without human GH 200 ng/ml. Relative abundance of IGF-I transcripts was evaluated by the RNase-protection assay, using a probe which permitted identification of initiation in exon 1 (site 1 (-380 bp from the 3' end of exon 1), site 2 (-343 bp), site 3 (-242 bp), sites 1 and 2 spliced, and site 4 (-32 bp)), as well as in exon 2. After normalization of signal intensity to adjust for differences in length of protected probe, the utilization of initiation sites in vitro was remarkably similar to that in vivo: 1, 14, 6, 23, 19 and 37% for sites 1, 2, 3, 1 and 2 spliced, 4 and exon 2 respectively in the cultured hepatocytes, compared with 1, ...Continue Reading

Citations

Oct 12, 2000·Molecular and Cellular Endocrinology·J L ZhuL S Phillips
Sep 13, 2000·Growth Hormone & IGF Research : Official Journal of the Growth Hormone Research Society and the International IGF Research Society·W W Lin, A M Oberbauer
Jul 18, 2001·The Journal of Biological Chemistry·E N KaytorL S Phillips
Sep 27, 2020·International Journal of Molecular Sciences·Aldona Kasprzak, Witold Szaflarski

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