Publication Date:
1987-07-17
Description:
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.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Lufkin, T -- Bancroft, C -- GM36847/GM/NIGMS NIH HHS/ -- New York, N.Y. -- Science. 1987 Jul 17;237(4812):283-6.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/3474782" target="_blank"〉PubMed〈/a〉
Keywords:
Acetyltransferases/genetics
;
Animals
;
Cell Fusion
;
Chloramphenicol O-Acetyltransferase
;
Gene Expression Regulation
;
Growth Hormone/genetics
;
Pituitary Gland/*physiology
;
Prolactin/*genetics
;
*Promoter Regions, Genetic
;
Rats
;
Transcription Factors/*genetics
;
Transcription, Genetic
;
Transfection
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
