Physiological and cellular responses caused by RNAi- mediated suppression of Snf7 orthologue in western corn rootworm (Diabrotica virgifera virgifera) larvae.
Ingestion of double stranded RNA (dsRNA) has been previously demonstrated to be effective in triggering RNA interference (RNAi) in western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte), providing potential novel opportunities for insect pest control. The putative Snf7 homolog of WCR (DvSnf7) has previously been shown to be an effective RNAi target for insect control, as DvSnf7 RNAi leads to lethality of WCR larvae. Snf7 functions as a part of the ESCRT (Endosomal Sorting Complex Required for Transport) pathway which plays a crucial role in cellular housekeeping by internalization, transport, sorting and lysosomal degradation of transmembrane proteins. To understand the effects that lead to death of WCR larvae by DvSnf7 RNAi, we examined some of the distinct cellular processes associated with ESCRT functions such as de-ubiquitination of proteins and autophagy. Our data indicate that ubiquitinated proteins accumulate in DvSnf7 dsRNA-fed larval tissues and that the autophagy process seems to be impaired. These findings suggest that the malfunctioning of these cellular processes in both midgut and fat body tissues triggered by DvSnf7 RNAi were the main effects leading to the death of WCR. This study also illustrates that Snf7 is an essential gene in WCR and its functions are consistent with biological functions described for other eukaryotes.
Department of Biotechnology, Monsanto Company, Chesterfield, Missouri, USA.
Parthasarathy
P
Ramaseshadri
Gerrit
G
Segers
Ronald
R
Flannagan
Elizabeth
E
Wiggins
William
W
Clinton
Oliver
O
Ilagan
Brian
B
McNulty
Thomas
T
Clark
Renata
R
Bolognesi
D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000502:physiology; D001343:Autophagy / Q000235:genetics / Q000502:physiology; D002468:Cell Physiological Phenomena; D001517:Coleoptera / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004064:Digestive System / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics* / Q000378:metabolism; D005216:Fat Body / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D007300:Insect Control / Q000379:methods; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D018613:Microscopy, Confocal; D008957:Models, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D057149:Ubiquitinated Proteins / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination; D003313:Zea mays / Q000469:parasitology
D056827:Endosomal Sorting Complexes Required for Transport; D019476:Insect Proteins; D034741:RNA, Small Interfering; D057149:Ubiquitinated Proteins; D049452:Green Fluorescent Proteins