JVI Accepts, published online ahead of print on 26 December 2007

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J. Virol. doi:10.1128/JVI.02060-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Sindbis virus genetic determinants of mosquito infection are associated with a highly conserved alphavirus and flavivirus envelope sequence

Dennis J. Pierro^*, Erik L. Powers, and Ken E. Olson

Arthropod-Borne and Infectious Disease Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523

^* To whom correspondence should be addressed. Email: pierrod{at}niaid.nih.gov.

The wild-type Sindbis virus (SINV) strain MRE16 efficiently infects Aedes aegypti midgut epithelial cells (MEC) but the laboratory derived neurovirulent SINV strain TE/5'2J poorly infects MEC. SINV determinants for MEC infection have been localized to the E2 glycoprotein. The E2 amino acid sequences of MRE16 and TE/5'2J differ at 60 residue sites. To identify the genetic determinants of MEC infection of MRE16, the TE/5'2J virus genome was altered to contain either domain chimeras or more focused nucleotide substitutions of MRE16. The growth patterns of derived viruses in cell culture were determined, as were the midgut infection rates (MIR) in Ae. aegypti mosquitoes. The results showed that substitution of the MRE16 E2 95-96 and the E2 116-119 amino acid residues into the TE/5'2J virus increase MIR both independently and in combination with each other. In addition, a unique PPF/.GDS aa motif was located between these two sites that was found to be a highly conserved sequence among alphaviruses and flaviviruses, but not other arboviruses.