Exploring the role of two interacting phosphoinositide 3-kinases of Haemonchus contortus.
BACKGROUND
Phosphoinositide 3-kinases (PI3Ks) are relatively conserved and important intracellular lipid kinases involved in signalling and other biological pathways. In the free-living nematode Caenorhabditis elegans, the heterodimeric form of PI3K consists of catalytic (AGE-1) and regulatory (AAP-1) subunits. These subunits are key components of the insulin-like signalling pathway and play roles in the regulation of the entry into and exit from dauer. Although, in parasitic nematodes, similar components are proposed to regulate the transition from free-living or arrested stages to parasitic larvae, nothing is known about PI3Ks in relation to the transition of third-stage larvae (L3s) to parasitism in Haemonchus contortus.


METHODS
An integrated molecular approach was used to investigate age-1 and aap-1 of H. contortus (Hc-age-1 and Hc-aap-1) in C. elegans.


RESULTS
The two genes Hc-age-1 and Hc-aap-1 were transcribed in all life stages, with the highest levels in the egg, infective L3 and adult female of H. contortus. The expression of these genes was localized to the intestine, contrasting the pattern of their orthologues in C. elegans (where they are expressed in both head neurons and the intestine). The yeast two-hybrid analysis demonstrated that the adaptor-binding domain of Hc-AGE-1 interacted strongly with the Hc-AAP-1; however, this complex did not rescue the function of its orthologue in age-1-deficient C. elegans.


CONCLUSIONS
This is the first time that the PI3K-encoding genes have been characterized from a strongylid parasitic nematode. The findings provide insights into the role of the PI3K heterodimer represented by Hc-age-1 and Hc-aap-1 in the developmental biology of H. contortus.
State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, 1 Shizishan Street, Wuhan, 430070, Hubei, China. li78561270@163.com.
Fa-Cai
FC
Li
Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Corner of Flemington Road and Park Drive, Parkville, VIC, 3010, Australia. robinbg@unimelb.edu.au.
Robin B
RB
Gasser
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 3800 Spruce Street, Philadelphia, PA19104, USA. jlok@vet.upenn.edu.
James B
JB
Lok
Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Corner of Flemington Road and Park Drive, Parkville, VIC, 3010, Australia. pasi.korhonen@unimelb.edu.au.
Pasi K
PK
Korhonen
State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, 1 Shizishan Street, Wuhan, 430070, Hubei, China. wangyifan_123456@163.com.
Yi-Fan
YF
Wang
State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, 1 Shizishan Street, Wuhan, 430070, Hubei, China. yinfangyuan96@126.com.
Fang-Yuan
FY
Yin
State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, 1 Shizishan Street, Wuhan, 430070, Hubei, China. 463771946@qq.com.
Li
L
He
State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, 1 Shizishan Street, Wuhan, 430070, Hubei, China. rzhou@mail.hzau.edu.cn.
Rui
R
Zhou
State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, 1 Shizishan Street, Wuhan, 430070, Hubei, China. zhaojunlong@mail.hzau.edu.cn.
Jun-Long
JL
Zhao
State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, 1 Shizishan Street, Wuhan, 430070, Hubei, China. mhu@mail.hzau.edu.cn.
Min
M
Hu
D000818:Animals; D005260:Female; D020869:Gene Expression Profiling; D006189:Haemonchus / Q000201:enzymology* / Q000254:growth & development; D007422:Intestines / Q000201:enzymology; D008018:Life Cycle Stages; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D025941:Protein Interaction Mapping; D021122:Protein Subunits / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D021122:Protein Subunits; D019869:Phosphatidylinositol 3-Kinases