Molecular characterization of the Haemonchus contortus phosphoinositide-dependent protein kinase-1 gene (Hc-pdk-1)

التفاصيل البيبلوغرافية
العنوان: Molecular characterization of the Haemonchus contortus phosphoinositide-dependent protein kinase-1 gene (Hc-pdk-1)
المؤلفون: Min Hu, Rui Zhou, Yanqin Zhou, Wenda Di, Li He, Fang-Yuan Yin, James B. Lok, Junlong Zhao, Robin B. Gasser, Facai Li, Pasi K. Korhonen
المصدر: Parasites & Vectors
بيانات النشر: BioMed Central, 2016.
سنة النشر: 2016
مصطلحات موضوعية: 0301 basic medicine, DNA, Complementary, 030231 tropical medicine, Molecular Sequence Data, Development, 3-Phosphoinositide-Dependent Protein Kinases, 03 medical and health sciences, 0302 clinical medicine, Haemonchus contortus, Gene expression, Daf-16, Animals, Protein kinase A, Caenorhabditis elegans, biology, Kinase, Research, Gene Expression Profiling, Sequence Analysis, DNA, biology.organism_classification, Cell biology, Gene expression profiling, Pleckstrin homology domain, 030104 developmental biology, Infectious Diseases, Immunology, Parasitology, Transgenesis, Haemonchus, pdk-1 gene
الوصف: Background Phosphoinositide-dependent protein kinase-1 (PDK-1), which functions downstream of phosphoinositide 3-kinase (AGE-1) and activates protein kinases of the AGC family, plays critical roles in regulating biology processes, such as metabolism, growth, development and survival. In the free-living nematode Caenorhabditis elegans, PDK-1 is a key component of the insulin-like signalling pathway, regulating the entry into and exit from dauer (arrested development). Although it is proposed that similar molecular mechanisms control the transition from the free-living to the parasitic stages of nematodes, nothing is known about PDK-1 in Haemonchus contortus, a socioeconomically important gastric nematode of ruminants. Methods Here, we isolated and characterized the pdk-1 gene (Hc-pdk-1) and its inferred product (Hc-PDK-1) from H. contortus. Using in vitro and in vivo methods, we then studied the transcriptional profiles of Hc-pdk-1 and anatomical gene expression patterns of Hc-PDK-1 in different developmental stages of C. elegans. Results In silico analysis of Hc-PDK-1 displayed conserved functional domains, such as protein kinase and pleckstrin homology (PH) domains and two predicted phosphorylation sites (Thr226/Tyr229), which are crucial for the phosphorylation of downstream signalling. The Hc-pdk-1 gene is transcribed in all of the main developmental stages of H. contortus, with its highest transcription in the infective third-stage larvae (iL3) compared with other stages. Transgene constructs, in which respective promoters were fused to the coding sequence for green fluorescent protein (GFP), were used to transform C. elegans, and to localize and compare the expression of Hc-pdk-1 and Ce-pdk-1. The expression of GFP under the control of the Hc-pdk-1 promoter was localized to the intestine, and head and tail neurons, contrasting somewhat the profile for the C. elegans ortholog, which is expressed in pharynx, intestine and head and tail neurons. Conclusions This is the first characterization of pdk-1/PDK-1 from a trichostrongyloid nematode. Taken together, the findings from this study provide a first glimpse of the involvement of Hc-pdk-1 in the insulin-like signalling pathway in H. contortus. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1351-6) contains supplementary material, which is available to authorized users.
اللغة: English
تدمد: 1756-3305
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e7df82d959282f92d44bc464929d75faTest
http://europepmc.org/articles/PMC4741024Test
حقوق: OPEN
رقم الانضمام: edsair.doi.dedup.....e7df82d959282f92d44bc464929d75fa
قاعدة البيانات: OpenAIRE