95e67fb14907c7995c9c46f720fd6729 pmc_33670223.pdf b4f9482d262fde13f7dc8540de0023480253c10c pmc_33670223.pdf 9dde2bed8b656a7b55f49024e07fa4644440a3e1224014b6529eadaa70891dab pmc_33670223.pdf Title: Time-Resolved Transcriptional Profiling of Epithelial Cells Infected by Intracellular Acinetobacter baumannii Subject: The rise in the number of antibiotic-resistant bacteria has become a serious threat to health, making it important to identify, characterize and optimize new molecules to help us to overcome the infections they cause. It is well known that Acinetobacter baumannii has a significant capacity to evade the actions of antibacterial drugs, leading to its emergence as one of the bacteria responsible for hospital and community-acquired infections. Nonetheless, how this pathogen infects and survives inside the host cell is unclear. In this study, we analyze the time-resolved transcriptional profile changes observed in human epithelial HeLa cells after infection by A. baumannii, demonstrating how it survives in host cells and starts to replicate 4 h post infection. These findings were achieved by sequencing RNA to obtain a set of Differentially Expressed Genes (DEGs) to understand how bacteria alter the host cells’ environment for their own benefit. We also determine common features observed in this set of genes and identify the protein–protein networks that reveal highly-interacted proteins. The combination of these findings paves the way for the discovery of new antimicrobial candidates for the treatment of multidrug-resistant bacteria. Keywords: Acinetobacter baumannii; infection; intracellular; transcriptional profile Author: Nuria Crua Asensio, Javier Macho Rendón and Marc Torrent Burgas Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Feb 26 01:24:15 2021 CET ModDate: Fri Feb 26 02:32:03 2021 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4105124 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- CEGHOZ+URWPalladioL-Ital Type 1 Custom yes yes no 10 0 BPFNAE+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 YHBUKN+URWPalladioL-BoldItal Type 1 Custom yes yes no 19 0 UXDIYQ+URWPalladioL-Roma Type 1 Custom yes yes no 23 0 SOZKOC+VnURWPalladioL-Bold Type 1 Custom yes yes no 27 0 MWHIBA+VnURWPalladioL Type 1 Custom yes yes no 32 0 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ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 12:09:04 CEST Size: 4105124 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 1043 FreeObjects: 171 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Bacterial Cell Culture Destination: subsection.2.1 Item: Title: Cell Culture and Infection Destination: subsection.2.2 Item: Title: Flow Cytometry Destination: subsection.2.3 Item: Title: Fluorescent Microscopy Destination: subsection.2.4 Item: Title: RNA Isolation Destination: subsection.2.5 Item: Title: Read Mapping and Differential Expression Analysis Destination: subsection.2.6 Item: Title: Clustering and Functional Enrichment Analysis Destination: subsection.2.7 Item: Title: Results and Discussion Destination: section.3 Children: Item: Title: Acinetobacter baumannii Can Survive inside Epithelial Cells Destination: subsection.3.1 Item: Title: Acinetobacter baumannii Reorganizes the Cell Transcriptome after Intracellular Invasion Destination: subsection.3.2 Item: Title: Regulation of Host Genes in Response to A. baumannii Infection Destination: subsection.3.3 Item: Title: Gene Expression Changes Are Correlated at the Level of Protein Protein Interactions Destination: subsection.3.4 Item: Title: Conclusions Destination: section.4 Item: Title: References Destination: figure.caption.7 Info: Title: Time-Resolved Transcriptional Profiling of Epithelial Cells Infected by Intracellular Acinetobacter baumannii Author: Nuria Crua Asensio, Javier Macho Rendón and Marc Torrent Burgas Subject: The rise in the number of antibiotic-resistant bacteria has become a serious threat to health, making it important to identify, characterize and optimize new molecules to help us to overcome the infections they cause. It is well known that Acinetobacter baumannii has a significant capacity to evade the actions of antibacterial drugs, leading to its emergence as one of the bacteria responsible for hospital and community-acquired infections. Nonetheless, how this pathogen infects and survives inside the host cell is unclear. In this study, we analyze the time-resolved transcriptional profile changes observed in human epithelial HeLa cells after infection by A. baumannii, demonstrating how it survives in host cells and starts to replicate 4 h post infection. These findings were achieved by sequencing RNA to obtain a set of Differentially Expressed Genes (DEGs) to understand how bacteria alter the host cells’ environment for their own benefit. We also determine common features observed in this set of genes and identify the protein protein networks that reveal highly-interacted proteins. The combination of these findings paves the way for the discovery of new antimicrobial candidates for the treatment of multidrug-resistant bacteria. 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