TY - JOUR
T1 - Juvenile hormone regulates Aedes aegypti Krüppel homolog 1 through a conserved E box motif
AU - Cui, Yingjun
AU - Sui, Yipeng
AU - Xu, Jingjing
AU - Zhu, Fang
AU - Palli, Subba Reddy
N1 - Funding Information:
This is contribution number 14-08-05 from the Kentucky Agricultural Experimental Station. This work was supported by National Institutes of Health ( GM070559-11 ).
PY - 2014/9
Y1 - 2014/9
N2 - Juvenile hormone (JH) plays important roles in regulation of many physiological processes including development, reproduction and metabolism in insects. However, the molecular mechanisms of JH signaling pathway are not completely understood. To elucidate the molecular mechanisms of JH regulation of Krüppel homolog 1 gene (Kr-h1) in Aedes aegypti, we employed JH-sensitive Aag-2cells developed from the embryos of this insect. In Aag-2cells, AaKr-h1 gene is induced by nanomolar concentration of JH III, its expression peaked at 1.5h after treatment with JH III. RNAi studies showed that JH induction of this gene requires the presence of Ae. aegypti methoprene-tolerant (AaMet). A conserved 13 nucleotide JH response element (JHRE, TGCCTC. CACGTGC) containing canonical E box motif (underlined) identified in the promoter of AaKr-h1 is required for JH induction of this gene. Critical nucleotides in the JHRE required for JH action were identified by employing mutagenesis and reporter assays. Reporter assays also showed that basic helix loop helix (bHLH) domain of AaMet is required for JH induction of AaKr-h1. 5' rapid amplification of cDNA ends method identified two isoforms of AaKr-h1, AaKr-h1α and AaKr-h1β, the expression of both isoforms is induced by JH III, but AaKr-h1α is the predominant isoform in both Aag-2cells and Ae. aegypti larvae.
AB - Juvenile hormone (JH) plays important roles in regulation of many physiological processes including development, reproduction and metabolism in insects. However, the molecular mechanisms of JH signaling pathway are not completely understood. To elucidate the molecular mechanisms of JH regulation of Krüppel homolog 1 gene (Kr-h1) in Aedes aegypti, we employed JH-sensitive Aag-2cells developed from the embryos of this insect. In Aag-2cells, AaKr-h1 gene is induced by nanomolar concentration of JH III, its expression peaked at 1.5h after treatment with JH III. RNAi studies showed that JH induction of this gene requires the presence of Ae. aegypti methoprene-tolerant (AaMet). A conserved 13 nucleotide JH response element (JHRE, TGCCTC. CACGTGC) containing canonical E box motif (underlined) identified in the promoter of AaKr-h1 is required for JH induction of this gene. Critical nucleotides in the JHRE required for JH action were identified by employing mutagenesis and reporter assays. Reporter assays also showed that basic helix loop helix (bHLH) domain of AaMet is required for JH induction of AaKr-h1. 5' rapid amplification of cDNA ends method identified two isoforms of AaKr-h1, AaKr-h1α and AaKr-h1β, the expression of both isoforms is induced by JH III, but AaKr-h1α is the predominant isoform in both Aag-2cells and Ae. aegypti larvae.
UR - http://www.scopus.com/inward/record.url?scp=84903783962&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84903783962&partnerID=8YFLogxK
U2 - 10.1016/j.ibmb.2014.05.009
DO - 10.1016/j.ibmb.2014.05.009
M3 - Article
C2 - 24931431
AN - SCOPUS:84903783962
SN - 0965-1748
VL - 52
SP - 23
EP - 32
JO - Insect Biochemistry and Molecular Biology
JF - Insect Biochemistry and Molecular Biology
IS - 1
ER -