Regulation of Rac1 and Reactive Oxygen Species Production in Response to Infection of Gastrointestinal Epithelia

Overview

Journal PLoS Pathog

Specialty Microbiology

Date 2016 Jan 14

PMID 26761793

Citations 36

Authors

Gerco Den Hartog

Ranajoy Chattopadhyay

Amber Ablack

Emily H Hall

Lindsay D Butcher

Asima Bhattacharyya

Lars Eckmann

Paul R Harris

Soumita Das

Peter B Ernst

Sheila E Crowe

Affiliations

Soon will be listed here.

Abstract

Generation of reactive oxygen species (ROS) during infection is an immediate host defense leading to microbial killing. APE1 is a multifunctional protein induced by ROS and after induction, protects against ROS-mediated DNA damage. Rac1 and NAPDH oxidase (Nox1) are important contributors of ROS generation following infection and associated with gastrointestinal epithelial injury. The purpose of this study was to determine if APE1 regulates the function of Rac1 and Nox1 during oxidative stress. Gastric or colonic epithelial cells (wild-type or with suppressed APE1) were infected with Helicobacter pylori or Salmonella enterica and assessed for Rac1 and NADPH oxidase-dependent superoxide production. Rac1 and APE1 interactions were measured by co-immunoprecipitation, confocal microscopy and proximity ligation assay (PLA) in cell lines or in biopsy specimens. Significantly greater levels of ROS were produced by APE1-deficient human gastric and colonic cell lines and primary gastric epithelial cells compared to control cells after infection with either gastric or enteric pathogens. H. pylori activated Rac1 and Nox1 in all cell types, but activation was higher in APE1 suppressed cells. APE1 overexpression decreased H. pylori-induced ROS generation, Rac1 activation, and Nox1 expression. We determined that the effects of APE1 were mediated through its N-terminal lysine residues interacting with Rac1, leading to inhibition of Nox1 expression and ROS generation. APE1 is a negative regulator of oxidative stress in the gastrointestinal epithelium during bacterial infection by modulating Rac1 and Nox1. Our results implicate APE1 in novel molecular interactions that regulate early stress responses elicited by microbial infections.

Citing Articles

Choroid plexus organoids reveal mechanisms of Streptococcus suis translocation at the blood-cerebrospinal fluid barrier.

Zhao T, Pellegrini L, van der Hee B, Boekhorst J, Fernandes A, Brugman S Fluids Barriers CNS. 2025; 22(1):14.

PMID: 39930492 PMC: 11812244. DOI: 10.1186/s12987-025-00627-y.

Helicobacter pylori and gastric cancer: mechanisms and new perspectives.

Duan Y, Xu Y, Dou Y, Xu D J Hematol Oncol. 2025; 18(1):10.

PMID: 39849657 PMC: 11756206. DOI: 10.1186/s13045-024-01654-2.

Alterations in Adipose Tissue and Adipokines in Heterozygous APE1/Ref-1 Deficient Mice.

Lee E, Jin H, Kim S, Joo H, Lee Y, An S Endocrinol Metab (Seoul). 2024; 39(6):932-945.

PMID: 39566547 PMC: 11695485. DOI: 10.3803/EnM.2024.2061.

Apurinic/apyrimidinic endodeoxyribonuclease 1 (APE1) promotes stress granule formation via YBX1 phosphorylation in ovarian cancer.

Mao S, Xie C, Liu Y, Zhao Y, Li M, Gao H Cell Mol Life Sci. 2024; 81(1):113.

PMID: 38436697 PMC: 10912283. DOI: 10.1007/s00018-023-05086-y.

Bacterial infection and microbiota in carcinogenesis and tumor development.

Li Q Front Cell Infect Microbiol. 2023; 13:1294082.

PMID: 38035341 PMC: 10684967. DOI: 10.3389/fcimb.2023.1294082.

References

Lambeth J . NOX enzymes and the biology of reactive oxygen. Nat Rev Immunol. 2004; 4(3):181-9. DOI: 10.1038/nri1312. View

Nardone G, Rocco A, Malfertheiner P . Review article: helicobacter pylori and molecular events in precancerous gastric lesions. Aliment Pharmacol Ther. 2004; 20(3):261-70. DOI: 10.1111/j.1365-2036.2004.02075.x. View

Ding S, OHara A, Denning T, Dirden-Kramer B, Mifflin R, Reyes V . Helicobacter pylori and H2O2 increase AP endonuclease-1/redox factor-1 expression in human gastric epithelial cells. Gastroenterology. 2004; 127(3):845-58. DOI: 10.1053/j.gastro.2004.06.017. View

Irani K, Xia Y, Zweier J, Sollott S, Der C, Fearon E . Mitogenic signaling mediated by oxidants in Ras-transformed fibroblasts. Science. 1997; 275(5306):1649-52. DOI: 10.1126/science.275.5306.1649. View

Nagata K, Yu H, Nishikawa M, KASHIBA M, Nakamura A, Sato E . Helicobacter pylori generates superoxide radicals and modulates nitric oxide metabolism. J Biol Chem. 1998; 273(23):14071-3. DOI: 10.1074/jbc.273.23.14071. View

Teshima S, Rokutan K, Nikawa T, Kishi K . Guinea pig gastric mucosal cells produce abundant superoxide anion through an NADPH oxidase-like system. Gastroenterology. 1998; 115(5):1186-96. DOI: 10.1016/s0016-5085(98)70090-3. View

Ernst P, Gold B . Helicobacter pylori in childhood: new insights into the immunopathogenesis of gastric disease and implications for managing infection in children. J Pediatr Gastroenterol Nutr. 1999; 28(5):462-73. DOI: 10.1097/00005176-199905000-00005. View

Kawahara T, Kohjima M, Kuwano Y, Mino H, Teshima-Kondo S, Takeya R . Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells. Am J Physiol Cell Physiol. 2004; 288(2):C450-7. DOI: 10.1152/ajpcell.00319.2004. View

DerMardirossian C, Bokoch G . GDIs: central regulatory molecules in Rho GTPase activation. Trends Cell Biol. 2005; 15(7):356-63. DOI: 10.1016/j.tcb.2005.05.001. View

10.

Dovas A, Couchman J . RhoGDI: multiple functions in the regulation of Rho family GTPase activities. Biochem J. 2005; 390(Pt 1):1-9. PMC: 1184558. DOI: 10.1042/BJ20050104. View

11.

Ernst P, Peura D, Crowe S . The translation of Helicobacter pylori basic research to patient care. Gastroenterology. 2006; 130(1):188-206. DOI: 10.1053/j.gastro.2005.06.032. View

12.

Cheng G, Diebold B, Hughes Y, Lambeth J . Nox1-dependent reactive oxygen generation is regulated by Rac1. J Biol Chem. 2006; 281(26):17718-26. DOI: 10.1074/jbc.M512751200. View

13.

Rokutan K, Kawahara T, Kuwano Y, Tominaga K, Sekiyama A, Teshima-Kondo S . NADPH oxidases in the gastrointestinal tract: a potential role of Nox1 in innate immune response and carcinogenesis. Antioxid Redox Signal. 2006; 8(9-10):1573-82. DOI: 10.1089/ars.2006.8.1573. View

14.

OHara A, Bhattacharyya A, Mifflin R, Smith M, Ryan K, Scott K . Interleukin-8 induction by Helicobacter pylori in gastric epithelial cells is dependent on apurinic/apyrimidinic endonuclease-1/redox factor-1. J Immunol. 2006; 177(11):7990-9. DOI: 10.4049/jimmunol.177.11.7990. View

15.

Bedard K, Krause K . The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiol Rev. 2007; 87(1):245-313. DOI: 10.1152/physrev.00044.2005. View

16.

Ding S, Minohara Y, Fan X, Wang J, Reyes V, Patel J . Helicobacter pylori infection induces oxidative stress and programmed cell death in human gastric epithelial cells. Infect Immun. 2007; 75(8):4030-9. PMC: 1952011. DOI: 10.1128/IAI.00172-07. View

17.

Guo Y, Chen J, Zhao T, Fan Z . Granzyme K degrades the redox/DNA repair enzyme Ape1 to trigger oxidative stress of target cells leading to cytotoxicity. Mol Immunol. 2008; 45(8):2225-35. DOI: 10.1016/j.molimm.2007.11.020. View

18.

Chattopadhyay R, Das S, Maiti A, Boldogh I, Xie J, Hazra T . Regulatory role of human AP-endonuclease (APE1/Ref-1) in YB-1-mediated activation of the multidrug resistance gene MDR1. Mol Cell Biol. 2008; 28(23):7066-80. PMC: 2593380. DOI: 10.1128/MCB.00244-08. View

19.

OHara A, Bhattacharyya A, Bai J, Mifflin R, Ernst P, Mitra S . Tumor necrosis factor (TNF)-alpha-induced IL-8 expression in gastric epithelial cells: role of reactive oxygen species and AP endonuclease-1/redox factor (Ref)-1. Cytokine. 2009; 46(3):359-69. PMC: 2846768. DOI: 10.1016/j.cyto.2009.03.010. View

20.

Bhattacharyya A, Chattopadhyay R, Burnette B, Cross J, Mitra S, Ernst P . Acetylation of apurinic/apyrimidinic endonuclease-1 regulates Helicobacter pylori-mediated gastric epithelial cell apoptosis. Gastroenterology. 2009; 136(7):2258-69. PMC: 2694750. DOI: 10.1053/j.gastro.2009.02.014. View