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Volume 270, Number 44, Issue of November 3, 1995 pp. 26570-26576
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Identification of Sequences Which Regulate the Expression of Drosophila melanogaster Doc Elements

(Received for publication, June 22, 1995; and in revised form, September 7, 1995)

Cristina Contursi Gabriella Minchiotti Pier Paolo Di Nocera

Long interspersed nuclear elements (LINEs) are mobile DNA elements which propagate by reverse transcription of RNA intermediates. LINEs lack long terminal repeats, and their expression is controlled by promoters located inside to the transcribed region of unit-length DNA copies. Doc elements constitute one of the seven families of LINEs found in Drosophila melanogaster. Plasmids in which the chloramphenicol acetyltransferase (CAT) gene is preceded by DNA segments from different Doc family members were used as templates for transient expression assays in Drosophila S2 cells. Transcription is initiated at the 5` end of Doc elements within hexamers fitting the consensus (C/G)AYTCG and is regulated by a DNA region which is located 20 base pairs (bp) downstream from the RNA start site(s). The region includes a sequence (RGACGTGY motif, or DE2) which stimulates transcription in other Drosophila LINEs, and two adjacent elements, DE1 and DE3. Moving the downstream region either 4 bp away from, or 5 bp closer to the RNA start site region inhibited transcription. Sequences located 200 bp downstream from the Doc 5` end repressed CAT expression in an orientation- and position-dependent manner. The inhibition reflects impaired translation of the CAT gene possibly consequent to the interaction of specific Doc RNA sequences with a cellular component.




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