Negative regulatory element associated with potentially functional promoter and enhancer elements in the long terminal repeats of endogenous murine leukemia virus-related proviral sequences
Three series of recombinant DNA clones were constructed, with the bacterial chloramphenical acetyltransferase (CAT) gene as a quantitative indicator, to examine the activities of promoter and enhancer sequence elements in the 5' long terminal repeat (LTR) of murine leukemia virus (MuLV)-related proviral sequences isolated from the mouse genome. Transient CAT expression was determined in mouse NIH 3T3, human HT1080, and mink CCL64 cultured cells transfected with the LTR-CAT constructs. The 700-base pair (bp) LTRs of three polytropic MuLV-related proviral clones and the 750-bp LTRs of four modified polytropic proviral clones, in complete structures either with or without the adjacent downstream sequences, all showed very little or negligible activities for CAT expression, while ecotropic MuLV LTRs were highly active. The MuLV-related LTRs were divided into three portions and examined separately. The 3' portion of the MuLV-related LTRs that contains the CCAAC and TATAA boxes was found to be a functional promoter, being about one-half to one-third as active as the corresponding portion of the ecotropic MuLV LTRs. A MboI-Bg/II fragment, representing the distinct 190- to 200-pb inserted segment in the middle, was found to be a potential enhancer, especially when examined in combination with the simian virus 40 promoter in CCL64 cells. A PstI-MboI fragment of the 5' portion, which contains the protein-binding motifs on the enhancer segment as well as the upstream LTF sequences, showed moderate enhancer activities in CCL6 cells but was virtually inactive in NIH 3T3 cells and HT1080 cells; addition of this fragment to the ecotropic LTR-CAT constructs depressed CAT expression.
- Research Organization:
- Oak Ridge National Lab., TN (USA)
- DOE Contract Number:
- AC05-84OR21400
- OSTI ID:
- 6156638
- Journal Information:
- J. Virol.; (United States), Vol. 63:6
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ANIMAL CELLS
GENE REGULATION
LEUKEMIA VIRUSES
DNA SEQUENCING
TRANSFERASES
GENES
DNA-CLONING
FIBROBLASTS
MAMMALS
MAN
MICE
RECOMBINANT DNA
SIMIAN VIRUS
TRANSCRIPTION
ANIMALS
CLONING
CONNECTIVE TISSUE CELLS
DNA
DNA HYBRIDIZATION
ENZYMES
HYBRIDIZATION
MICROORGANISMS
NUCLEIC ACIDS
ONCOGENIC VIRUSES
ORGANIC COMPOUNDS
PARASITES
PRIMATES
RODENTS
SOMATIC CELLS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
VIRUSES
550200* - Biochemistry
550900 - Pathology