thap and atf-2 regulated sterol carrier protein-2 promoter activities in the larval midgut of the yellow fever mosquito, aedes aegyptithap和atf-2监管甾醇载体protein-2子活动黄热病蚊子的幼虫中肠,埃及伊蚊.pdfVIP

THAP and ATF-2 Regulated Sterol Carrier Protein-2 Promoter Activities in the Larval Midgut of the Yellow Fever Mosquito, Aedes aegypti 1 2 1 2 2 Rong Peng *, Qiang Fu , Huazhu Hong , Tyler Schwaegler , Que Lan * 1 College of Life Sciences, Central China Normal University, Wuhan, Hubei, China, 2 Department of Entomology, University of Wisconsin, Madison, Wisconsin, United States of America Abstract Expression of sterol carrier protein-2 (SCP-2) in Aedes aegypti shows a distinct temporal/spatial pattern throughout the life cycle. In order to identify the transcription factors responsible for the larval temporal/spatial regulation of AeSCP-2 transcription, AeSCP-2 promoter activities were studied in vivo via transient transfection of promoter/reporter gene assays. Regulatory sequences upstream 21.3 kb of the transcription start site of AeSCP-2 were found to be critical for the in vivo temporal/spatial promoter activity. Interestingly, the 21.6 kb promoter sequence efficiently drove the larval midgut-specific siRNA expression, indicating that the 21.6 kb upstream sequence is sufficient for temporal/spatial AeSCP-2 transcriptional activity. Four transcription factors were identified in the midgut nuclear extract from feeding larvae via labeled 21.6/ 21.3 kb DNA probe pull-down and proteomic analysis. Co-transfection of the promoter/reporter gene with inducible siRNA expression of each transcription factor was performed to confirm the regulatory function of individual transcription factor on AeSCP-2 transcriptional activities in the larval midgut. The results indicate that two of the identified transcription factors, Thanatos-associated protein (THAP) and activating transcription factor-2 (ATF-2),