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©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
HP0953 - hypothetical virulence factor overexpresion and localization during Helicobacter pylori infection of gastric epithelium
Nancy K Arteaga-Resendiz, Gerardo E Rodea, Rosa María Ribas-Aparicio, Alma L Olivares-Cervantes, Juan Arturo Castelán-Vega, José de Jesús Olivares-Trejo, Sandra Mendoza-Elizalde, Edgar O López-Villegas, Christian Colín, Pamela Aguilar-Rodea, Alfonso Reyes-López, Marcela Salazar García, Norma Velázquez-Guadarrama
Nancy K Arteaga-Resendiz, Gerardo E Rodea, Alma L Olivares-Cervantes, Sandra Mendoza-Elizalde, Christian Colín, Pamela Aguilar-Rodea, Norma Velázquez-Guadarrama, Laboratorio de Investigación en Enfermedades Infecciosas, Hospital Infantil de México Federico Gómez, Mexico City 06720, Mexico
Nancy K Arteaga-Resendiz, Rosa María Ribas-Aparicio, Juan Arturo Castelán-Vega, 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
José de Jesús Olivares-Trejo, Laboratorio de Adquisición de Hierro, Universidad Autónoma de la Ciudad México, Posgrado Ciencias Genómica, Mexico City 03100, Mexico
Edgar O López-Villegas, Laboratorio Central de Microscopía, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City 11340, Mexico
Alfonso Reyes-López, Centro de estudios económicos y sociales en salud, Hospital Infantil de México Federico Gómez, Mexico City 06720, Mexico
Marcela Salazar García, Laboratorio de Investigación en Biología del Desarrollo y Teratogénesis Experimental, Hospital Infantil de México Federico Gómez, Mexico City 06720, Mexico
Author contributions: Arteaga-Resendiz NK, Rodea GE, Olivares-Cervantes AL, and Colín C performed the experiments, and acquired and analyzed data; Arteaga-Resendiz NK, Rodea GE, Colín C, Olivares-Cervantes AL, Aguilar-Rodea P, Ribas-Aparicio RM, López-Villegas EO, Olivares-Trejo JJ, and Velázquez-Guadarrama N interpreted the data; Reyes-López A and Arteaga-Resendiz NK statistical data analyzed; Rodea GE, Ribas-Aparicio RM, Mendoza-Elizalde S, Salazar García M and Velázquez-Guadarrama N wrote the manuscript; and all authors approved the final version of the article.
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; and the Instituto Politécnico Nacional by Consejo Nacional de Ciencia y Tecnología, CB-222180.
Institutional review board statement: The study was reviewed and approved by the Ethics, Biosafety and Scientific committees at the Health Institute approved the experiment (HIM/2009/037. SSA851 and HIM/2014/012. SSA 1098).
Institutional animal care and use committee statement: Animal care was performed under national and institutional policies (NOM-062-ZOO-1999).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to them.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Norma Velázquez-Guadarrama, PhD, Research Scientist, Laboratorio de Investigación en Enfermedades Infecciosas, Hospital Infantil de México Federico Gómez, Dr. Márquez 162, Col. Doctores, Alcaldía Cuauhtémoc, Mexico City 06720, Mexico. normave@himfg.edu.mx
Received: January 29, 2022
Peer-review started: January 29, 2022
First decision: April 10, 2022
Revised: April 26, 2022
Accepted: July 11, 2022
Article in press: July 11, 2022
Published online: August 7, 2022
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 340 hypothetical 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.
Core Tip: The high prevalence and persistence of Helicobacter pylori infection and the diverse pathologies associated with it suggest that the virulence factors of this microorganism are varied. Moreover, its proteome contains 340 hypothetical proteins, so it is crucial to investigate them to elucidate the mechanisms of its virulence and survival. We studied the hypothetical protein HP0953, its location around infected cells, and the probable posttranslational modifications. Its similarity to an exotoxin suggests that HP0953 is a virulence factor.
