title: Expression profiling and comparative analyses of seven midgut serine proteases from the yellow fever mosquito, Aedes aegypti.
abstract: Aedes aegypti utilizes blood for energy production, egg maturation and replenishment of maternal reserves. The principle midgut enzymes responsible for bloodmeal digestion are endoproteolytic serine-type proteases within the S1.A subfamily. While there are hundreds of serine protease-like genes in the A. aegypti genome, only five are known to be expressed in the midgut. We describe the cloning, sequencing and expression profiling of seven additional serine proteases and provide a genomic and phylogenetic assessment of these findings. Of the seven genes, four are constitutively expressed and three are transcriptionally induced upon blood feeding. The amount of transcriptional induction is strongly correlated among these genes. Alignments reveal that, in general, the conserved catalytic triad, active site and accessory catalytic residues are maintained in these genes and phylogenetic analysis shows that these genes fall within three distinct clades; trypsins, chymotrypsins and serine collagenases. Interestingly, a previously described trypsin consistently arose with other serine collagenases in phylogenetic analyses. These results suggest that multiple gene duplications have arisen within the S1.A subfamily of midgut serine proteases and/or that A. aegypti has evolved an array of proteases with a broad range of substrate specificities for rapid, efficient digestion of bloodmeals.
authors.affiliation: Arthropod Infectious Disease Laboratory, 1692 Campus Delivery, Department of Microbiology, Immunology and Pathology at Colorado State University, Fort Collins, CO 80523, USA. dbrackney@salud.unm.edu
authors.forename: Doug E
authors.initials: DE
authors.lastname: Brackney
authors.forename: Jun
authors.initials: J
authors.lastname: Isoe
authors.forename: Black
authors.initials: B
authors.lastname: W C
authors.forename: Jorge
authors.initials: J
authors.lastname: Zamora
authors.forename: Brian D
authors.initials: BD
authors.lastname: Foy
authors.forename: Roger L
authors.initials: RL
authors.lastname: Miesfeld
authors.forename: Ken E
authors.initials: KE
authors.lastname: Olson
mesh_terms: D000330:Aedes / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D004064:Digestive System / Q000737:chemistry / Q000201:enzymology; D020869:Gene Expression Profiling; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D057057:Serine Proteases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D057057:Serine Proteases