Eaton, Lynne Tracey; (1991) Transcriptional regulation of retrovirus-like VL30 genetic elements. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

The nucleotide sequence of the long terminal repeats (LTRs) of the retrovirus - transmissible mouse VL30 cDNA clones, NVL-1 and NVL-2, were determined and compared with that of the prototype clone NVL-3. The three LTRs shared a typical U3 R U5 structure and showed the unusual features of redundancy in the tRNAgly primer binding site and adjacent inverted repeat. The NVL-1 and NVL-2 LTRs were almost identical and differed from the NVL-3 LTR in the U3 domain harbouring transcriptional regulatory determinants. The VL30 elements were serum responsive and showed elevated levels of expression in many transformed cell lines. SI nuclease analysis showed that all the NVL VL30 elements responded to cellular transformation, but only NVL-1/2 elements were serum responsive. Both types of response were mediated via protein kinase C-independent pathways. Transient expression assays showed that the NVL-1/2 and NVL-3 U3 domains possess promoter and enhancer activities. The U3 region of both types of LTR contained enhancer determinants important in controlling the response to N-ras as well as sequences important for controlling basal levels of expression. However the U3 domain of the NVL-1/2 LTRs did not confer serum responsiveness. These results showed that the U3 domains of the NVL VL30 LTRs contain cis acting regulatory determinants important in controlling basal levels of expression and the response to cellular transformation. However the mechanisms controlling the serum response are at least in part dissociable from those responsible for cell transformation regulated expression.

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