HP0953-hypothetical virulence factor overexpresion and localization during Helicobacter pylori infection of gastric epithelium

查看全文 作　　者：Nancy K Arteaga-[1,2]Resendiz;Gerardo E [1]Rodea;Rosa María Ribas-[2]Aparicio;Alma L Olivares-[1]Cervantes;Juan Arturo Castelán-[2]Vega;Joséde Jesús Olivares-[3]Trejo;Sandra Mendoza-[1]Elizalde;Edgar O López-[4]Villegas;Christian [1]Colín;Pamela Aguilar-[1]Rodea;Alfonso Reyes-[5]López;Marcela Salazar [6]García;Norma Velázquez-[1]Guadarrama

机构地区：[1]Laboratorio de Investigación en Enfermedades Infecciosas,Hospital Infantil de México Federico Gómez,Mexico City 06720,Mexico;[2]Posgrado en Biomedicina y Biotecnología Molecular,Laboratorio de Producción y Control de Biológicos,Escuela Nacional de Ciencias Biológicas,Instituto Politécnico Nacional,Mexico City 11340,Mexico;[3]Laboratorio de Adquisición de Hierro,Universidad Autónoma de la Ciudad México,Posgrado Ciencias Genómica,Mexico City 03100,Mexico;[4]Laboratorio Central de Microscopía,Escuela Nacional de Ciencias Biológicas,Instituto Politécnico Nacional,Mexico City 11340,Mexico;[5]Centro de estudios económicos y sociales en salud,Hospital Infantil de México Federico Gómez,Mexico City 06720,Mexico;[6]Laboratorio de Investigación en Biología del Desarrollo y Teratogénesis Experimental,Hospital Infantil de México Federico Gómez,Mexico City 06720,Mexico

出　　处：《World Journal of Gastroenterology》索引2022年第28卷第29期,共17页

基　　金：Supported by the Federal Funds,HIM/2009/037.SSA851 and HIM/2014/012.SSA 1098;the grant from Secretaría de Investigación y Posgrado,SIP 20161878;the Instituto Politécnico Nacional by Consejo Nacional de Ciencia y Tecnología,CB-222180. 摘　　要：BACKGROUND The high prevalence and persistence of Helicobacter pylori(H. pylori) infection, as well as the diversity of pathologies related to it, suggest that the virulence factors used by this microorganism are varied. Moreover, as its proteome contains 340hypothetical proteins, it is important to investigate them to completely understand the mechanisms of its virulence and survival. We have previously reported that the hypothetical protein HP0953 is overexpressed during the first hours of adhesion to inert surfaces, under stress conditions, suggesting its role in the environmental survival of this bacterium and perhaps as a virulence factor.AIM To investigate the expression and localization of HP0953 during adhesion to an inert surface and against gastric(AGS) cells.METHODS Expression analysis was performed for HP0953 during H. pylori adhesion. HP0953 expression at 0,3, 12, 24, and 48 h was evaluated and compared using the Kruskal-Wallis equality-of-populations rank test. Recombinant protein was produced and used to obtain polyclonal antibodies for immunolocalization. Immunogold technique was performed on bacterial sections during adherence to inert surfaces and AGS cells, which was analyzed by transmission electron microscopy. HP0953 protein sequence was analyzed to predict the presence of a signal peptide and transmembrane helices, both provided by the ExPASy platform, and using the GLYCOPP platform for glycosylation sites. Different programs, via, I-TASSER, RaptorX, and HHalign-Kbest, were used to perform three-dimensional modeling.RESULTS HP0953 exhibited its maximum expression at 12 h of infection in gastric epithelium cells.Immunogold technique revealed HP0953 localization in the cytoplasm and accumulation in some peripheral areas of the bacterial body, with greater expression when it is close to AGS cells.Bioinformatics analysis revealed the presence of a signal peptide that interacts with the transmembrane region and then allows the release of the protein to the external environment. The programs also showed a similarity with the Tip-alpha protein of H. pylori. Tip-alpha is an exotoxin that penetrates cells and induces tumor necrosis factor alpha production, and HP0953 could have a similar function as posttranslational modification sites were found;modifications in turn require enzymes located in eukaryotic cells. Thus, to be functional, HP0953 may necessarily need to be translocated inside the cell where it can trigger different mechanisms producing cellular damage.CONCLUSION The location of HP0953 around infected cells, the probable posttranslational modifications, and its similarity to an exotoxin suggest that this protein is a virulence factor. 关键词：Hypothetical protein HP0953 ADHERENCE Helicobacter pylori GLYCOCALYX Virulence factor PERSISTENCE

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HP0953-hypothetical virulence factor overexpresion and localization during Helicobacter pylori infection of gastric epithelium

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