Protein crosslinking by transglutaminase controls cuticle morphogenesis in Drosophila.
Transglutaminase (TG) plays important and diverse roles in mammals, such as blood coagulation and formation of the skin barrier, by catalyzing protein crosslinking. In invertebrates, TG is known to be involved in immobilization of invading pathogens at sites of injury. Here we demonstrate that Drosophila TG is an important enzyme for cuticle morphogenesis. Although TG activity was undetectable before the second instar larval stage, it dramatically increased in the third instar larval stage. RNA interference (RNAi) of the TG gene caused a pupal semi-lethal phenotype and abnormal morphology. Furthermore, TG-RNAi flies showed a significantly shorter life span than their counterparts, and approximately 90% of flies died within 30 days after eclosion. Stage-specific TG-RNAi before the third instar larval stage resulted in cuticle abnormality, but the TG-RNAi after the late pupal stage did not, indicating that TG plays a key role at or before the early pupal stage. Immediately following eclosion, acid-extractable protein from wild-type wings was nearly all converted to non-extractable protein due to wing maturation, whereas several proteins remained acid-extractable in the mature wings of TG-RNAi flies. We identified four proteins--two cuticular chitin-binding proteins, larval serum protein 2, and a putative C-type lectin-as TG substrates. RNAi of their corresponding genes caused a lethal phenotype or cuticle abnormality. Our results indicate that TG-dependent protein crosslinking in Drosophila plays a key role in cuticle morphogenesis and sclerotization.
Graduate School of Systems Life Sciences, Kyushu University, Fukuoka, Japan.
Toshio
T
Shibata
Shigeru
S
Ariki
Naoaki
N
Shinzawa
Ryuta
R
Miyaji
Haruka
H
Suyama
Miyuki
M
Sako
Nobuyuki
N
Inomata
Takumi
T
Koshiba
Hirotaka
H
Kanuka
Shun-Ichiro
S
Kawabata
D000818:Animals; D004330:Drosophila / Q000254:growth & development*; D029721:Drosophila Proteins / Q000378:metabolism*; D007814:Larva / Q000254:growth & development*; D009024:Morphogenesis; D010641:Phenotype; D034622:RNA Interference; D013379:Substrate Specificity; D011503:Transglutaminases / Q000235:genetics / Q000378:metabolism*
D029721:Drosophila Proteins; D011503:Transglutaminases