title: Role of Penaeus monodon hemocyte homeostasis associated protein (PmHHAP) in regulation of caspase-mediated apoptosis.
abstract: The viral responsive protein, PmHHAP, plays an important role in the control of hemocyte homeostasis in shrimps during viral infection. In this study, we further investigate the role of PmHHAP in the regulation of hemocyte apoptosis. RNA interference (RNAi) mediated gene silencing was used to suppress the PmHHAP expression and the change in hemocyte apoptosis was determined in the knockdown shrimp. Within circulating hemocytes, PmHHAP knockdown increased the number of annexin V-positive apoptotic cells and the combined caspase-3/-7 activity and induced the characteristic apoptotic DNA ladder. Furthermore, PmHHAP down-regulation was accompanied by significantly altered expression of apoptosis-related proteins including the effector caspases, PmCaspase and PmCasp. Yeast two-hybrid and co-immunoprecipitation assays showed that PmHHAP binds to the p20 domain of PmCasp. Moreover, the recombinant PmHHAP protein was able to reduce the caspase activity in the actinomycin D-treated hemocyte cells and rPmCasp-treated hemocyte cells. Taken together, our data indicate that PmHHAP regulates hemocyte homeostasis by inhibits apoptotic cell death through caspase activation.
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, 254 Phayathai Road, Bangkok 10330, Thailand.
authors.forename: Kantamas
authors.initials: K
authors.lastname: Apitanyasai
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, 254 Phayathai Road, Bangkok 10330, Thailand; National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand.
authors.forename: Piti
authors.initials: P
authors.lastname: Amparyup
authors.affiliation: National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand; Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Mahidol University, Rama VI Road, Bangkok 10400, Thailand.
authors.forename: Walaiporn
authors.initials: W
authors.lastname: Charoensapsri
authors.affiliation: National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand; Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Mahidol University, Rama VI Road, Bangkok 10400, Thailand.
authors.forename: Saengchan
authors.initials: S
authors.lastname: Senapin
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, 254 Phayathai Road, Bangkok 10330, Thailand. Electronic address: anchalee.k@chula.ac.th.
authors.forename: Anchalee
authors.initials: A
authors.lastname: Tassanakajon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000276:immunology*; D060829:Arthropod Proteins / Q000235:genetics / Q000276:immunology*; D053148:Caspase 3 / Q000276:immunology*; D053179:Caspase 7 / Q000276:immunology*; D006434:Hemocytes / Q000276:immunology; D033561:Penaeidae / Q000235:genetics / Q000276:immunology*; D034622:RNA Interference; D034741:RNA, Small Interfering; D016415:Sequence Alignment
chemical_list: D051017:Apoptosis Regulatory Proteins; D060829:Arthropod Proteins; D034741:RNA, Small Interfering; D053148:Caspase 3; D053179:Caspase 7
keywords: Apoptosis; Caspase; Hemocyte homeostasis; Penaeus monodon; Shrimp immunity