This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Park, J H Articles by Taylor, M W Search for Related Content PubMed PubMed Citation Articles by Park, J H Articles by Taylor, M W

Analysis of signals controlling expression of the Chinese hamster ovary aprt gene.

Department of Biology, Indiana University, Bloomington 47405.

ABSTRACT

The 5' end of the Chinese hamster ovary aprt gene was sequenced and transcription start sites were determined by both S1 nuclease protection and primer extension assays. Deletion mutants covering the same area were constructed, and adenine phosphoribosyltransferase (APRT) or chloramphenicol acetyltransferase (CAT) activity was measured by transient-expression assays. The aprt gene uses a single cluster of transcription start sites and lacks consensus sequences such as TATA and CCAAT, which are general components of eucaryotic promoters. The 5' deletion mutations of the promoter sequences demonstrated that (i) there is no decrease in either APRT activity or transcription extending to position -89 (relative to the main transcription start site); (ii) an additional 29-base-pair (bp) deletion decreases APRT activity and transcription twofold; and (iii) a deletion past the transcription start sites (P5' delta +27) abolishes both APRT activity and transcription, indicating that a 60-bp fragment immediately upstream of the main transcription start site is involved in basic transcription and a 29-bp fragment just upstream of the 60 bp-fragment stimulates transcription twofold. The 3' deletion mutations showed that a deletion of a 61-bp fragment in the 5' leader and coding sequence abolishes the efficient translation of an aprt-CAT gene transcript. In addition, there are two polyadenylation signals at the genomic 3' end, with the proximal one being sufficient for functional polyadenylation.

This article has been cited by other articles:

• Rody, A, Holtrich, U, Solbach, C, Kourtis, K, von Minckwitz, G, Engels, K, Kissler, S, Gatje, R, Karn, T, Kaufmann, M (2005). Methylation of estrogen receptor {beta} promoter correlates with loss of ER-{beta} expression in mammary carcinoma and is an early indication marker in premalignant lesions. Endocr Relat Cancer 12: 903-916 [Abstract] [Full Text]  
• Alfonzo, J. D., Crother, T. R., Guetsova, M. L., Daignan-Fornier, B., Taylor, M. W. (1999). APT1, but Not APT2, Codes for a Functional Adenine Phosphoribosyltransferase in Saccharomyces cerevisiae. J. Bacteriol. 181: 347-352 [Abstract] [Full Text]  
• Paulin, R. P., Ho, T., Balzer, H. J., Holliday, R. (1998). Gene Silencing by DNA Methylation and Dual Inheritance in Chinese Hamster Ovary Cells. Genetics 149: 1081-1088 [Abstract] [Full Text]  
• Hewitt, S. M., Fraizer, G. C., Saunders, G. F. (1995). Transcriptional Silencer of the Wilms' Tumor Gene WT1 Contains an Alu Repeat. J. Biol. Chem. 270: 17908-17912 [Abstract] [Full Text]