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1: Gene 1996 Oct 10;175(1-2):193-7

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Cloning and upstream sequence of a juvenile hormone-regulated gene from the migratory locust.

Zhang J, Wyatt GR

Department of Biology, Queen's University, Kingston, Ont, Canada.

As a step toward analyzing the molecular mechanism of action of juvenile hormone (JH), the gene encoding a JH-inducible 21-kDa protein (Jhp21) produced in the fat body of the migratory locust has been cloned. Four exons, representing 750 nucleotides of cDNA sequence, were found to be distributed through 13 kb of genomic DNA. Upstream 2 kb of DNA has been sequenced and three potential hormone-response elements have been identified.

PMID: 8917098, UI: 97074671

2: Mol Cell Endocrinol 1996 Aug 30;122(1):15-20

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Juvenile hormone regulation of an insect gene: a specific transcription factor and a DNA response element.

Zhang J, Saleh DS, Wyatt GR

Department of Biology, Queen's University, Kingston, Ontario, Canada.

We have used locust fat body nuclear protein extracts and upstream DNA of the juvenile hormone (JH)-inducible locust gene, jhp21, to examine the regulation of specific transcription by JH. Promoter activity was assayed with G-free cassette reporter constructs. Nuclear extracts from adult female fat body, previously exposed to JH or an analog, actively transcribe from the jhp21 promoter and a control adenovirus major late (AdML) promoter, whereas extracts from JH-deprived female fat body, or other tissues, transcribe strongly from the AdML promoter but weakly or not at all from the jhp21 promoter. Transcription is enhanced by sequences between -140 and -211 nt from the jhp21 transcription start point (tsp), which include a CAAT box, and also by sequences between -1056 and -1200. A 15-nt partially palindromic sequence element found at -1152, resembling known hormone response elements, was shown to stimulate transcription when restored to truncated jhp21 DNA. Two very similar sequences occur further upstream. In electrophoretic mobility shift assays (EMSA), the same sequence element was shown to specifically bind a protein that was present in nuclear extracts from JH-exposed, but not from JH-deprived, fat body. Several lines of evidence suggest that the DNA element may be a JH response element (JHRE). The JH-induced protein that binds to it appears to be a transcription factor that activates the initiation of JH target gene (jhp21) transcription, and could be a JH receptor.

PMID: 8898344, UI: 97053965