Deletion mapping of tumor promotion–susceptibility gene pro1 implicates an RNA polymerase III transcription unit
✍ Scribed by Robert R. Garrity; Glenn A. Hegamyer; John L. Seed; Nancy H. Colburn
- Publisher
- John Wiley and Sons
- Year
- 2006
- Tongue
- English
- Weight
- 738 KB
- Volume
- 3
- Category
- Article
- ISSN
- 0899-1987
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✦ Synopsis
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.