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Transposon mutagenesis

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https://www.readbyqxmd.com/read/28724549/identification-of-new-tumor-suppressor-genes-in-triple-negative-breast-cancer
#1
Roberto Rangel, Liliana Guzman-Rojas, Takahiro Kodama, Michiko Kodama, Justin Y Newberg, Neal G Copeland, Nancy A Jenkins
Although genomic sequencing has provided a better understating of the genetic landmarks in triple-negative breast cancer (TNBC), functional validation of candidate cancer genes (CCG) remains unsolved. In this study, we used a transposon mutagenesis strategy based on a two-step Sleeping Beauty (SB) forward genetic screen to identify and validate new tumor suppressors (TS) in this disease. We generated 120 siRNAs targeting 40 SB-identified candidate breast cancer TS genes and used them to downregulate expression of these genes in four human TNBC cell lines...
July 19, 2017: Cancer Research
https://www.readbyqxmd.com/read/28698237/maize-yabby-genes-drooping-leaf1-and-drooping-leaf2-regulate-plant-architecture
#2
Josh Strable, Jason G Wallace, Erica Unger-Wallace, Sarah Briggs, Peter Bradbury, Edward S Buckler, Erik Vollbrecht
Leaf architecture directly influences canopy structure, consequentially affecting yield. We discovered a maize (Zea mays) mutant with aberrant leaf architecture, which we named drooping leaf1 (drl1). Pleiotropic mutations in drl1 affect leaf length and width, leaf angle, and internode length and diameter. These phenotypes are enhanced by natural variation at the drl2 enhancer locus, including reduced expression of the drl2-Mo17 allele in the Mo17 inbred. A second drl2 allele, produced by transposon mutagenesis, interacted synergistically with drl1 mutants and reduced drl2 transcript levels...
July 11, 2017: Plant Cell
https://www.readbyqxmd.com/read/28683522/role-of-eptc-in-biofilm-formation-by-campylobacter-jejuni-nctc11168-on-polystyrene-and-glass-surfaces
#3
Eun Seob Lim, Joo-Sung Kim
The complex roles of cell surface modification in the biofilm formation of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are poorly understood. In a screen of mutants from random transposon mutagenesis, an insertional mutation in eptC gene (cj0256) resulted in a significant decrease in C. jejuni NCTC11168 biofilm formation (<20%) on major food contact surfaces, such as polystyrene and borosilicate glass, compared with wild-type cells (p < 0.05). In C. jejuni strain 81-176, the protein encoded by eptC modifies cell surface structures, such as lipid A, the inner core of lipooligosaccharide, and the flagellar rod protein (FlgG), by attaching phosphoethanolamine...
July 7, 2017: Journal of Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28680437/knots-untie-molecular-determinants-involved-in-knot-formation-induced-by-pseudomonas-savastanoi-in-woody-hosts
#4
REVIEW
Eloy Caballo-Ponce, Jesús Murillo, Marta Martínez-Gil, Alba Moreno-Pérez, Adrián Pintado, Cayo Ramos
The study of the molecular basis of tree diseases is lately receiving a renewed attention, especially with the emerging perception that pathogens require specific pathogenicity and virulence factors to successfully colonize woody hosts. Pathosystems involving woody plants are notoriously difficult to study, although the use of model bacterial strains together with genetically homogeneous micropropagated plant material is providing a significant impetus to our understanding of the molecular determinants leading to disease...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28650484/insertional-mutagenesis-identifies-drivers-of-a-novel-oncogenic-pathway-in-invasive-lobular-breast-carcinoma
#5
Sjors M Kas, Julian R de Ruiter, Koen Schipper, Stefano Annunziato, Eva Schut, Sjoerd Klarenbeek, Anne Paulien Drenth, Eline van der Burg, Christiaan Klijn, Jelle J Ten Hoeve, David J Adams, Marco J Koudijs, Jelle Wesseling, Micha Nethe, Lodewyk F A Wessels, Jos Jonkers
Invasive lobular carcinoma (ILC) is the second most common breast cancer subtype and accounts for 8-14% of all cases. Although the majority of human ILCs are characterized by the functional loss of E-cadherin (encoded by CDH1), inactivation of Cdh1 does not predispose mice to develop mammary tumors, implying that mutations in additional genes are required for ILC formation in mice. To identify these genes, we performed an insertional mutagenesis screen using the Sleeping Beauty transposon system in mice with mammary-specific inactivation of Cdh1...
June 26, 2017: Nature Genetics
https://www.readbyqxmd.com/read/28614737/glucose-triggers-the-cytotoxicity-of-citrobacter-sp-r1-against-microcystis-aeruginosa
#6
Pengfei Sun, Sofia Esquivel-Elizondo, Yuhua Zhao, Yonghong Wu
Algicidal bacteria offer a promising option for killing Microcystis aeruginosa, one notorious cyanobacteria causing harmful algal blooms. In this study, Citrobacter sp. R1 presented high algicidal activity (81.6±2.2%, 72h) against M. aeruginosa when cultured using glucose, while it showed no algicidal activity (0±3.4%) when cultured using wheat bran, suggesting that appropriate carbon source is crucial for algicidal bacteria in killing M. aeruginosa. The underlying algicidal mechanism of strain R1 was explored by studying the effect of different carbon sources (glucose and wheat bran) on its key algicidal gene expression and total protein translation...
June 11, 2017: Science of the Total Environment
https://www.readbyqxmd.com/read/28614382/genome-wide-transposon-mutagenesis-of-proteus-mirabilis-essential-genes-fitness-factors-for-catheter-associated-urinary-tract-infection-and-the-impact-of-polymicrobial-infection-on-fitness-requirements
#7
Chelsie E Armbruster, Valerie Forsyth-DeOrnellas, Alexandra O Johnson, Sara N Smith, Lili Zhao, Weisheng Wu, Harry L T Mobley
The Gram-negative bacterium Proteus mirabilis is a leading cause of catheter-associated urinary tract infections (CAUTIs), which are often polymicrobial. Numerous prior studies have uncovered virulence factors for P. mirabilis pathogenicity in a murine model of ascending UTI, but little is known concerning pathogenesis during CAUTI or polymicrobial infection. In this study, we utilized five pools of 10,000 transposon mutants each and transposon insertion-site sequencing (Tn-Seq) to identify the full arsenal of P...
June 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28584132/transposon-mutagenesis-identifies-chromatin-modifiers-cooperating-with-ras-in-thyroid-tumorigenesis-and-detects-atxn7-as-a-cancer-gene
#8
Cristina Montero-Conde, Luis J Leandro-Garcia, Xu Chen, Gisele Oler, Sergio Ruiz-Llorente, Mabel Ryder, Iñigo Landa, Francisco Sanchez-Vega, Konnor La, Ronald A Ghossein, Dean F Bajorin, Jeffrey A Knauf, Jesse D Riordan, Adam J Dupuy, James A Fagin
Oncogenic RAS mutations are present in 15-30% of thyroid carcinomas. Endogenous expression of mutant Ras is insufficient to initiate thyroid tumorigenesis in murine models, indicating that additional genetic alterations are required. We used Sleeping Beauty (SB) transposon mutagenesis to identify events that cooperate with Hras(G12V) in thyroid tumor development. Random genomic integration of SB transposons primarily generated loss-of-function events that significantly increased thyroid tumor penetrance in Tpo-Cre/homozygous FR-Hras(G12V) mice...
June 20, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28575524/identifying-transposon-insertions-and-their-effects-from-rna-sequencing-data
#9
Julian R de Ruiter, Sjors M Kas, Eva Schut, David J Adams, Marco J Koudijs, Lodewyk F A Wessels, Jos Jonkers
Insertional mutagenesis using engineered transposons is a potent forward genetic screening technique used to identify cancer genes in mouse model systems. In the analysis of these screens, transposon insertion sites are typically identified by targeted DNA-sequencing and subsequently assigned to predicted target genes using heuristics. As such, these approaches provide no direct evidence that insertions actually affect their predicted targets or how transcripts of these genes are affected. To address this, we developed IM-Fusion, an approach that identifies insertion sites from gene-transposon fusions in standard single- and paired-end RNA-sequencing data...
June 1, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28557013/unravelling-the-complexity-and-redundancy-of-carbon-catabolic-repression-in-the-plant-growth-promoting-bacterium-pseudomonas-fluorescens-sbw25
#10
Yunhao Liu, Chaitanya S Gokhale, Paul B Rainey, Xue-Xian Zhang
The two-component system CbrAB is the principal regulator for cellular metabolic balance in Pseudomonas fluorescens SBW25 and is necessary for growth on many substrates including xylose. To understand the regulatory linkage between CbrAB and genes for xylose utilization (xut), we performed transposon mutagenesis analysis of ΔcbrB to select for Xut(+) suppressors. This led to identification of crc and hfq. Subsequent genetic and biochemical analysis showed that Crc and Hfq are key mediators of succinate-provoked carbon catabolite repression (CCR)...
May 29, 2017: Molecular Microbiology
https://www.readbyqxmd.com/read/28550060/surface-sensing-for-flagellar-gene-expression-on-solid-media-in-paenibacillus-sp-naist15-1
#11
Kazuo Kobayashi, Yu Kanesaki, Hirofumi Yoshikawa
A rhizosphere Gram-positive bacterial isolate, Paenibacillus sp. NAIST15-1, exhibits intriguing motility behavior on hard agar media. Paenibacillus sp. shows increased transcription of flagellar genes and hyperflagellation when transferred from liquid to solid media. Hyperflagellated cells form wandering colonies that are capable of moving around on the surface of media containing ≥1.5% agar. Transposon mutagenesis was used to identify genes critical for motility. In addition to flagellar genes, this mutagenesis identified five non-flagellar structural genes that were important for motility...
May 26, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28515302/transposon-mutagenesis-of-the-zika-virus-genome-highlights-regions-essential-for-rna-replication-and-restricted-for-immune-evasion
#12
Benjamin O Fulton, David Sachs, Megan C Schwarz, Peter Palese, Matthew J Evans
The molecular constraints affecting Zika virus (ZIKV) evolution are not well understood. To investigate ZIKV genetic flexibility, we used transposon mutagenesis to add 15-nucleotide insertions throughout the MR766 ZIKV genome and subsequently deep sequenced the viable mutants. Few ZIKV insertion mutants replicated, which likely reflects a high degree of functional constraints on the genome. The NS1 gene exhibited distinct mutational tolerances at different stages of the screen. This result may define regions of the NS1 protein that are required for the different stages of the viral life cycle...
May 17, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28512090/the-vibrio-cholerae-vexgh-rnd-efflux-system-maintains-cellular-homeostasis-by-effluxing-vibriobactin
#13
Dillon E Kunkle, X Renee Bina, James E Bina
Resistance-nodulation-division (RND) superfamily efflux systems have been widely studied for their role in antibiotic resistance, but their native biological functions remain poorly understood. We previously showed that loss of RND-mediated efflux in Vibrio cholerae resulted in activation of the Cpx two-component regulatory system, which mediates adaptation to stress resulting from misfolded membrane proteins. Here, we investigated the mechanism linking RND-mediated efflux to the Cpx response. We performed transposon mutagenesis screening of RND-deficient V...
May 16, 2017: MBio
https://www.readbyqxmd.com/read/28509393/nmea-a-novel-efflux-transporter-specific-for-nucleobases-and-nucleosides-contributes-to-metal-resistance-in-aspergillus-nidulans
#14
Sofia Balaska, Vassilios Myrianthopoulos, Martha Tselika, Dimitris G Hatzinikolaou, Emmanuel Mikros, George Diallinas
Through Minos transposon mutagenesis we obtained A. nidulans mutants resistant to 5-fluorouracil due to insertions into the upstream region of the uncharacterized gene nmeA, encoding a Major Facilitator Superfamily (MFS) transporter. Minos transpositions increased nmeA transcription, which is otherwise extremely low under all conditions tested. To dissect the function of NmeA we used strains overexpressing or genetically lacking the nmeA gene. Strains overexpressing NmeA are resistant to toxic purine analogues, but also, to cadmium, zinc and borate, whereas an isogenic nmeAΔ null mutant exhibits increased sensitivity to these compounds...
May 16, 2017: Molecular Microbiology
https://www.readbyqxmd.com/read/28507068/genome-wide-survey-of-pseudomonas-aeruginosa-pa14-reveals-a-role-for-the-glyoxylate-pathway-and-extracellular-proteases-in-the-utilization-of-mucin
#15
Jeffrey M Flynn, Chi Phan, Ryan C Hunter
Chronic airway infections by the opportunistic pathogen Pseudomonas aeruginosa are a major cause of mortality in cystic fibrosis (CF) patients. While this bacterium has been extensively studied for its virulence determinants, biofilm growth and immune evasion mechanisms, comparatively little is known about the nutrient sources that sustain its growth in vivo Respiratory mucins represent a potentially abundant bioavailable nutrient source, though we have recently shown that canonical pathogens inefficiently use these host glycoproteins as a growth substrate...
May 15, 2017: Infection and Immunity
https://www.readbyqxmd.com/read/28500044/rpon-%C3%AF-54-is-required-for-floc-formation-but-not-for-extracellular-polysaccharide-biosynthesis-in-a-floc-forming-aquincola-tertiaricarbonis-strain
#16
Dianzhen Yu, Ming Xia, Liping Zhang, Yulong Song, You Duan, Tong Yuan, Minjie Yao, Liyou Wu, Chunyuan Tian, Zhenbin Wu, Xiangzhen Li, Jizhong Zhou, Dongru Qiu
Some bacteria are capable of forming flocs, in which bacterial cells become self-flocculated by secreted extracellular polysaccharides and other biopolymers. The floc-forming bacteria play a central role in activated sludge, which has been widely utilized for the treatment of municipal sewage and industrial wastewater. Here, we use a floc-forming bacterium, Aquincolatertiaricarbonis RN12, as a model to explore the biosynthesis of extracellular polysaccharides and the regulation of floc formation. A large gene cluster for exopolysaccharide biosynthesis and a gene encoding the alternative sigma factor RpoN1, one of the four paralogues, have been identified in floc formation-deficient mutants generated by transposon mutagenesis, and the gene functions have been further confirmed by genetic complementation analyses...
July 15, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28499399/identification-of-potential-campylobacter-jejuni-genes-involved-in-biofilm-formation-by-ez-tn5-transposome-mutagenesis
#17
Amy Huei Teen Teh, Sui Mae Lee, Gary A Dykes
BACKGROUND: Biofilm formation has been suggested to play a role in the survival of Campylobacter jejuni in the environment and contribute to the high incidence of human campylobacteriosis. Molecular studies of biofilm formation by Campylobacter are sparse. RESULTS: We attempted to identify genes that may be involved in biofilm formation in seven C. jejuni strains through construction of mutants using the EZ-Tn5 Transposome system. Only 14 mutants with reduced biofilm formation were obtained, all from one strain of C...
May 12, 2017: BMC Research Notes
https://www.readbyqxmd.com/read/28489862/novel-genes-associated-with-enhanced-motility-of-escherichia-coli-st131
#18
Asha Kakkanat, Minh-Duy Phan, Alvin W Lo, Scott A Beatson, Mark A Schembri
Uropathogenic Escherichia coli (UPEC) is the cause of ~75% of all urinary tract infections (UTIs) and is increasingly associated with multidrug resistance. This includes UPEC strains from the recently emerged and globally disseminated sequence type 131 (ST131), which is now the dominant fluoroquinolone-resistant UPEC clone worldwide. Most ST131 strains are motile and produce H4-type flagella. Here, we applied a combination of saturated Tn5 mutagenesis and transposon directed insertion site sequencing (TraDIS) as a high throughput genetic screen and identified 30 genes associated with enhanced motility of the reference ST131 strain EC958...
2017: PloS One
https://www.readbyqxmd.com/read/28487432/deciphering-mcr-2-colistin-resistance
#19
Jian Sun, Yongchang Xu, Rongsui Gao, Jingxia Lin, Wenhui Wei, Swaminath Srinivas, Defeng Li, Run-Shi Yang, Xing-Ping Li, Xiao-Ping Liao, Ya-Hong Liu, Youjun Feng
Antibiotic resistance is a prevalent problem in public health worldwide. In general, the carbapenem β-lactam antibiotics are considered a final resort against lethal infections by multidrug-resistant bacteria. Colistin is a cationic polypeptide antibiotic and acts as the last line of defense for treatment of carbapenem-resistant bacteria. Very recently, a new plasmid-borne colistin resistance gene, mcr-2, was revealed soon after the discovery of the paradigm gene mcr-1, which has disseminated globally. However, the molecular mechanisms for MCR-2 colistin resistance are poorly understood...
May 9, 2017: MBio
https://www.readbyqxmd.com/read/28483441/the-cytidine-repressor-participates-in-the-regulatory-pathway-of-indole-in-pantoea-agglomerans
#20
Mengqi Jia, Xuemei Yu, Jing Jiang, Zihua Li, Yongjun Feng
Indole, an important signal molecule in both intraspecies and interspecies, regulates a variety of bacterial behaviors, but its regulatory mechanism is still unknown. Pantoea agglomerans YS19, a preponderant endophytic bacterium isolated from rice, does not produce indole, yet it senses exogenous indole. In this study, a mutant of YS19-Rp(r) whose target gene expression was downregulated by indole was selected through mTn5 transposon mutagenesis. Using the TAIL-PCR technique, the mutation gene was identified as a cytR homologue, which encodes a cytidine repressor (CytR) protein, a bacterial transcription factor involved in a complex regulation scheme...
May 5, 2017: Research in Microbiology
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