title: Neisseria meningitidis causes cell cycle arrest of human brain microvascular endothelial cells at S phase via p21 and cyclin G2.
abstract: Microbial pathogens have developed several mechanisms to modulate and interfere with host cell cycle progression. In this study, we analysed the effect of the human pathogen Neisseria meningitidis on cell cycle in a brain endothelial cell line as well as in primary brain endothelial cells. We found that N.  Meningitidis causes an accumulation of cells in the S phase early at 3 and at 24 h post-infection that was paralleled by a decrease of cells in G2/M phase. Importantly, the outer membrane proteins of the colony opacity-associated (Opa) protein family as well as the Opc protein proved to trigger the accumulation of cells in the S phase. A focused cell cycle reverse transcription quantitative polymerase chain reaction-based array and integrated network analysis revealed changes in the abundance of several cell cycle regulatory mRNAs, including the cell cycle inhibitors p21(WAF1/CIP1) and cyclin G2. These alterations were reflected in changes in protein expression levels and/or relocalization in N. meningitidis-infected cells. Moreover, an increase in p21(WAF1/CIP1) expression was found to be p53 independent. Genetic ablation of p21(WAF1/CIP1) and cyclin G2 abrogated N. meningitidis-induced S phase accumulation. Finally, by measuring the levels of the biomarker 8-hydroxydeoxyguanosine and phosphorylation of the histone variant H2AX, we provide evidence that N. meningitidis induces oxidative DNA damage in infected cells.
authors.affiliation: Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Wilhelm F
authors.initials: WF
authors.lastname: Oosthuysen
authors.affiliation: Department of Bioinformatics, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Mueller
authors.affiliation: Department of Bioinformatics, University of Wuerzburg, Wuerzburg, Germany.|Institute of Human Genetics, University of Wuerzburg, Germany.
authors.forename: Marcus T
authors.initials: MT
authors.lastname: Dittrich
authors.affiliation: Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Schubert-Unkmeir
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001921:Brain; D059447:Cell Cycle Checkpoints; D002478:Cells, Cultured; D056767:Cyclin G2 / Q000378:metabolism*; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism*; D042783:Endothelial Cells / Q000382:microbiology* / Q000502:physiology; D020869:Gene Expression Profiling; D054884:Host-Pathogen Interactions; D006801:Humans; D009345:Neisseria meningitidis / Q000472:pathogenicity*; D060888:Real-Time Polymerase Chain Reaction; D016196:S Phase
chemical_list: D001425:Bacterial Outer Membrane Proteins; C493956:CDKN1A protein, human; D056767:Cyclin G2; D050759:Cyclin-Dependent Kinase Inhibitor p21; C074517:opc protein, bacteria; C090348:Opa protein, Neisseria