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Deletion mapping of tumor promotion–susceptibility gene pro 1 implicates an RNA polymerase III transcription unit
Author(s) -
Garrity Robert R.,
Hegamyer Glenn A.,
Seed John L.,
Colburn Nancy H.
Publication year - 1990
Publication title -
molecular carcinogenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.254
H-Index - 97
eISSN - 1098-2744
pISSN - 0899-1987
DOI - 10.1002/mc.2940030412
Subject(s) - biology , microbiology and biotechnology , rna , transcription (linguistics) , rna polymerase ii , gene , promoter , gene expression , genetics , philosophy , linguistics
Abstract The murine gene pro 1 has been cloned from JB6 epidermal cell lines that are sensitive to neoplastic transformation by tumor promoters. Insensitive JB6 variants acquire susceptibility to neoplastic transformation by tumor promoters when transfected with pro 1. The repetitive nature of pro 1 was indicated by sequence and Southern analysis. In contrast, northern analysis of RNA from promotion‐sensitive cells revealed the presence of a small pro 1‐hybridizing transcript. Strand‐specific RNA probes implicated an RNA polymerase III (RNAPIII) coding domain in pro 1 as the source of this hybridization signal. Ribonuclease protection of gel‐purified pro 1 RNA from JB6 variant cell lines identified a 130‐nucleotide transcript. The size of this transcript is compatible with in vitro RNAPIII transcription of pro 1. Deletion mapping of pro 1 by exonuclease III demonstrated that the biologically active domain included the RNAPIII transcription unit. RNA probes map pro 1 RNA within the activity domain. These results delineate an activity domain of 597 nucleotides and suggest that a small RNA is the product of promotion‐sensitivity gene pro 1.