Identification by cell fusion of gene sequences that interact with positive trans-acting factors

doi:10.1126/science.3474782

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Science 17 July 1987:
Vol. 237. no. 4812, pp. 283 - 286
DOI: 10.1126/science.3474782

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Science, Vol 237, Issue 4812, 283-286
Copyright © 1987 by American Association for the Advancement of Science

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Identification by cell fusion of gene sequences that interact with positive trans-acting factors

T Lufkin and C Bancroft

Cell fusion experiments have implicated positive or negative regulatory factors in the cell type--specific expression of specialized endogenous genes. The inability to readily manipulate such genes has prevented characterization of the cis-acting DNA sequences that interact with these factors. A transfection-fusion technique, which combined stable gene transfer and formation of transient heterokaryons, was used to study this class of factors and their DNA binding sites. Messenger RNA directed by a quiescent, rat prolactin promoter region stably transferred into mouse fibroblasts was detected only after fusion to rat pituitary cells, implying that pituitary cells contain a positive cell type--specific factor or factors. Nuclear run-on assays showed that fusion activation is transcriptional. Fusion did not activate either a stably transferred rat growth hormone gene promoter or expression of the endogenous silent fibroblast prolactin or growth hormone genes. Analysis by 5'-deletion mutation identified a 30-base pair DNA sequence required for cell fusion activation of the rat prolactin promoter region. Comparison with previous results from direct cellular transfer of this region implies that transfection-fusion identifies novel regulatory DNA sequences.

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