COPI-mediated blood meal digestion in vector mosquitoes is independent of midgut ARF-GEF and ARF-GAP regulatory activities

Jun Isoe, Weston Stover, R. Barrett Miesfeld, Roger Miesfeld

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We have previously shown that defects in COPI coatomer proteins cause 80% mortality in blood fed Aedes aegypti mosquitoes by 96h post-feeding. In this study we show that similar deficiencies in COPII and clathrin mediated vesicle transport do not disrupt blood meal digestion and are not lethal, even though both COPII and clathrin functions are required for ovarian development. Since COPI vesicle transport is controlled in mammalian cells by upstream G proteins and associated regulatory factors, we investigated the function of the orthologous ADP-ribosylation factor 1 (ARF1) and ARF4 proteins in mosquito tissues. We found that both ARF1 and ARF4 function upstream of COPI vesicle transport in blood fed mosquitoes given that an ARF1/ARF4 double deficiency is required to phenocopy the feeding-induced mortality observed in COPI coatomer deficient mosquitoes. Small molecule inhibitors of guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs) are often transitory, and therefore, we investigated the role of five Ae. aegypti ARF-GEF and ARF-GAP proteins in blood meal digestion using RNA interference. Surprisingly, we found that ARF-GEF and ARF-GAP functions are not required for blood meal digestion, even though both vitellogenesis and ovarian development in Ae. aegypti are dependent on GBF1 (ARF-GEF) and GAP1/GAP2 (ARF-GAPs) proteins. Moreover, deficiencies in orthologous COPI regulating genes in Anopheles stephensi mosquitoes had similar phenotypes, indicating conserved functions in these two mosquito species. We propose that based on the need for rapid initiation of protein digestion and peritrophic membrane formation, COPI vesicle transport in midgut epithelial cells is not dependent on ARF-GEF and ARF-GAP regulatory proteins to mediate vesicle recycling within the first 48h post-feeding.

Original languageEnglish (US)
Pages (from-to)732-739
Number of pages8
JournalInsect Biochemistry and Molecular Biology
Volume43
Issue number8
DOIs
StatePublished - Aug 2013

Fingerprint

Coat Protein Complex I
Guanine Nucleotide Exchange Factors
blood meal
Culicidae
midgut
Meals
Digestion
Transport Vesicles
ADP-Ribosylation Factor 1
Blood
digestion
Aedes aegypti
clathrin
Clathrin
ovarian development
Proteins
proteins
Coatomer Protein
Vitellogenesis
GTPase-activating proteins

Keywords

  • Mortality
  • Secretion
  • Vesicle transport
  • Vitellogenesis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Insect Science

Cite this

COPI-mediated blood meal digestion in vector mosquitoes is independent of midgut ARF-GEF and ARF-GAP regulatory activities. / Isoe, Jun; Stover, Weston; Miesfeld, R. Barrett; Miesfeld, Roger.

In: Insect Biochemistry and Molecular Biology, Vol. 43, No. 8, 08.2013, p. 732-739.

Research output: Contribution to journalArticle

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