A cell-free mRNA stability assay reveals conservation of the enzymes and mechanisms of mRNA decay between mosquito and mammalian cell lines.
The rate of mRNA turnover is an important determinant of levels of gene expression. Although this process has been studied extensively in mammalian cells and yeast, relatively little is known about the mRNA decay pathways in insects. Our analysis found that the vast majority of components of the mRNA decay machinery are conserved between humans and mosquitoes. Moreover, the half-lives of Aedes albopictus mRNAs are within a similar range to those of mammalian mRNAs. In order to investigate mechanistic aspects of mRNA decay in mosquitoes, we developed an in vitro system using cytoplasmic S100 extracts from A. albopictus C6/36 cells. Using this decay assay, we show here that all the pathways of mRNA turnover that have been observed in mammalian cells (deadenylation, decapping, 3'-to-5' exonucleolytic decay and 5'-to-3' exonucleolytic decay) are active in C6/36 extracts. Finally, we present compelling evidence that the major deadenylase in C6/36 extracts is likely to be a homolog of the human poly(A) specific ribonuclease, PARN. Our results suggest a high level of conservation in the factors and pathways of mRNA decay between mosquitoes and humans.
Opyrchal
Mateusz
M
Department of Microbiology, Immunology & Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, 80523, USA.
Anderson
John R
JR
Sokoloski
Kevin J
KJ
Wilusz
Carol J
CJ
Wilusz
Jeffrey
J
D000330:Aedes; D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles; D001483:Base Sequence; D002460:Cell Line; D017124:Conserved Sequence; D003465:Culex; D009033:Culicidae / Q000378:metabolism*; D017931:DNA Primers; D003609:Dactinomycin / Q000494:pharmacology; D004355:Drug Stability; D007700:Kinetics; D008322:Mammals; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D012333:RNA, Messenger; D003609:Dactinomycin