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J. Biol. Chem., Vol. 279, Issue 5, 3308-3317, January 30, 2004

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Characterization of Two Mosquito STATs, AaSTAT and CtSTAT

DIFFERENTIAL REGULATION OF TYROSINE PHOSPHORYLATION AND DNA BINDING ACTIVITY BY LIPOPOLYSACCHARIDE TREATMENT AND BY JAPANESE ENCEPHALITIS VIRUS INFECTION^*

Chang-Chi Lin, Chih-Ming Chou¶, Ya-Li Hsu||, Jih-Ching Lien, Yu-Ming Wang, Shui-Tsung Chen**, Shu-Chuan Tsai**, Pei-Wen Hsiao, and Chang-Jen Huang**

From the Graduate Institute of Life Science and Institute of Preventive Medicine, National Defense Medical Center, Taipei 114, the ^¶Department of Biochemistry, Taipei Medical University, Taipei 117, and the Institutes of ^||Zoology, ^**Biological Chemistry, and BioAgricultural Sciences, Academia Sinica, Taipei 115, Taiwan

Two mosquito STATs, AaSTAT and CtSTAT, have been cloned from Aedes albopictus and Culex tritaeniorhynchus mosquitoes, respectively. These two STATs are more similar to those of Drosophila, Anopheles, and mammalian STAT5 in the DNA binding and Src homology 2 domains. The mRNA transcripts are expressed at all developmental stages, and the proteins are present predominantly at the pupal and adult stages in both mosquitoes. Stimulation with lipopolysaccharide resulted in an increase of tyrosine phosphorylation and DNA binding activity of AaSTAT and CtSTAT as well as an increase of luciferase activity of a reporter gene containing Drosophila STAT binding motif in mosquito C6/36 cells. After being infected with Japanese encephalitis virus, nuclear extracts of C6/36 cells revealed a decrease of tyrosine phosphorylation and DNA binding activity of AaSTAT which could be restored by sodium orthovanadate treatment. Taking all of the data together, this is the first report to clone and characterize two mosquito STATs with 81% identity and to demonstrate a different response of tyrosine phosphorylation and DNA binding of these two STATs by lipopolysaccharide treatment and by Japanese encephalitis virus infection.

Received for publication, September 3, 2003 , and in revised form, November 5, 2003.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s)AY299686 and AY299687.

To whom correspondence should be addressed: Institute of Biological Chemistry, Academia Sinica, 128, Sec 2, Academia Rd., Taipei 115, Taiwan. Tel.: 886-2-2785-5696; Fax: 886-2-2788-9759; E-mail: cjibc{at}gate.sinica.edu.tw.

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