title: Mycoplasma gallisepticum (HS strain) surface lipoprotein pMGA interacts with host apolipoprotein A-I during infection in chicken.
abstract: The adhesin protein from Mycoplasma gallisepticum (HS strain), namely pMGA1.2, is required for M. gallisepticum (MG) infection in chicken. However, the host factor(s) that interact with pMGA1.2 is not known. In this study, we prepared the membrane fraction of trachea epithelial cells from chicken embryos. Using an improved virus overlay protein blot assay (VOPBA) and glutathione S-transferase (GST) pull-down assay, we found that pMGA1.2 specifically bound to a ∼30 kDa host protein. This host protein was further identified by mass spectrometry as chicken apolipoprotein A-I (ApoA-I). We expressed and purified the recombinant ApoA-I protein in Escherichia coli and confirmed that it bound to the purified pMGA1.2 protein in vitro. Transiently expressed pMGA1.2 and ApoA-I were colocalized in HeLa cells. Finally, we designed small interfering RNA (siRNA) molecules to knock down the expression of either ApoA-I or pMGA1.2, which inhibited the MG-induced cell cycle disruption in cells of chicken embryo fibroblast cell line (DF-1). Similarly, knockdown of ApoA-I inhibited the cilia loss and damage in chicken trachea cells in MG infection. In summary, ApoA-I may be an essential host factor in MG infection through interacting with pMGA1.2.
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Fuli
authors.initials: F
authors.lastname: Hu
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Chengcheng
authors.initials: C
authors.lastname: Zhao
authors.affiliation: China National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Dingren
authors.initials: D
authors.lastname: Bi
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Tian
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Jiao
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Biological Sciences and Border Biomedical Research Center, University of Texas at El Paso, El Paso, TX, 79968, USA. jsun@utep.edu.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Sun
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, China. xlpengsishun@mail.hzau.edu.cn.|China National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China. xlpengsishun@mail.hzau.edu.cn.
authors.forename: Xiuli
authors.initials: X
authors.lastname: Peng
mesh_terms: D000818:Animals; D016632:Apolipoprotein A-I / Q000235:genetics / Q000378:metabolism*; D030161:Avian Proteins / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D054884:Host-Pathogen Interactions; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D009175:Mycoplasma Infections / Q000235:genetics / Q000378:metabolism / Q000382:microbiology / Q000662:veterinary*; D045686:Mycoplasma gallisepticum / Q000235:genetics / Q000378:metabolism*; D011201:Poultry Diseases / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D011485:Protein Binding
chemical_list: D016632:Apolipoprotein A-I; D030161:Avian Proteins; D001426:Bacterial Proteins; D008074:Lipoproteins
keywords: Apolipoprotein A-I; Chicken; Mycoplasma gallisepticum HS strain; pMGA1.2