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Non-B Dna-Forming Motifs Promote Mfd-Dependent Stationary-Phase Mutagenesis In Bacillus Subtilis, Tatiana Ermi, Carmen Vallin, Ana Gabriela Regalado García, Moises Bravo, Ismaray Fernandez Cordero, Holly Anne Martin, Mario Pedraza-Reyes, Eduardo Robleto Jun 2021

Non-B Dna-Forming Motifs Promote Mfd-Dependent Stationary-Phase Mutagenesis In Bacillus Subtilis, Tatiana Ermi, Carmen Vallin, Ana Gabriela Regalado García, Moises Bravo, Ismaray Fernandez Cordero, Holly Anne Martin, Mario Pedraza-Reyes, Eduardo Robleto

Life Sciences Faculty Research

Transcription-induced mutagenic mechanisms limit genetic changes to times when expression happens and to coding DNA. It has been hypothesized that intrinsic sequences that have the potential to form alternate DNA structures, such as non-B DNA structures, influence these mechanisms. Non-B DNA structures are promoted by transcription and induce genome instability in eukaryotic cells, but their impact in bacterial genomes is less known. Here, we investigated if G4 DNA-and hairpin-forming motifs influence stationary-phase mutagenesis in Bacillus subtilis. We developed a system to measure the influence of non-B DNA on B. subtilis stationary-phase mutagenesis by deleting the wild-type argF at its chromosomal …


Genomics, Exometabolomics, And Metabolic Probing Reveal Conserved Proteolytic Metabolism Of Thermoflexus Hugenholtzii And Three Candidate Species From China And Japan, Scott C. Thomas, Devon Payne, Kevin O. Tamadonfar, Cale O. Seymour, Jian Yu Jiao, Senthil K. Murugapiran, Dengxun Lai, Rebecca Lau, Benjamin P. Bowen, Leslie P. Silva, Katherine B. Louie, Marcel Huntemann, Alicia Clum, Alex Spunde, Manoj Pillay, Krishnaveni Palaniappan, Neha Varghese, Natalia Mikhailova, I. Min Chen, Dimitrios Stamatis, T. B.K. Reddy, Ronan O’Malley, Chris Daum, Nicole Shapiro, Natalia Ivanova, Nikos C. Kyrpides, Tanja Woyke, Emiley Eloe-Fadrosh, Trinity L. Hamilton, Paul Dijkstra, Brian P. Hedlund May 2021

Genomics, Exometabolomics, And Metabolic Probing Reveal Conserved Proteolytic Metabolism Of Thermoflexus Hugenholtzii And Three Candidate Species From China And Japan, Scott C. Thomas, Devon Payne, Kevin O. Tamadonfar, Cale O. Seymour, Jian Yu Jiao, Senthil K. Murugapiran, Dengxun Lai, Rebecca Lau, Benjamin P. Bowen, Leslie P. Silva, Katherine B. Louie, Marcel Huntemann, Alicia Clum, Alex Spunde, Manoj Pillay, Krishnaveni Palaniappan, Neha Varghese, Natalia Mikhailova, I. Min Chen, Dimitrios Stamatis, T. B.K. Reddy, Ronan O’Malley, Chris Daum, Nicole Shapiro, Natalia Ivanova, Nikos C. Kyrpides, Tanja Woyke, Emiley Eloe-Fadrosh, Trinity L. Hamilton, Paul Dijkstra, Brian P. Hedlund

Life Sciences Faculty Research

Thermoflexus hugenholtzii JAD2 , the only cultured representative of the Chloroflexota order Thermoflexales, is abundant in Great Boiling Spring (GBS), NV, United States, and close relatives inhabit geothermal systems globally. However, no defined medium exists for T. hugenholtzii JAD2 and no single carbon source is known to support its growth, leaving key knowledge gaps in its metabolism and nutritional needs. Here, we report comparative genomic analysis of the draft genome of T. hugenholtzii JAD2 and eight closely related metagenome-assembled genomes (MAGs) from geothermal sites in China, Japan, and the United States, representing “Candidatus Thermoflexus japonica,” “Candidatus Thermoflexus tengchongensis,” and “Candidatus …


Bacterial Cyclic Diguanylate Signaling Networks Sense Temperature, Henrik Almblad, Trevor E. Randall, Fanny Liu, Katherine Leblanc, Ryan A. Groves, Weerayuth Kittichotirat, Geoffrey L. Winsor, Nicolas Fournier, Emily Au, Julie Groizeleau, Jacquelyn D. Rich, Yuefei Lou, Elise Granton, Laura K. Jennings, Larissa A. Singletary, Tara M.L. Winstone, Nathan M. Good, Roger E. Bumgarner, Michael F. Hynes, Manu Singh, Maria Silvina Stietz, Fiona S.L. Brinkman, Ayush Kumar, Ann Karen Cornelia Brassinga, Matthew R. Parsek, Boo Shan Tseng, Ian A. Lewis, Bryan G. Yipp, Justin L. Maccallum, Joe Jonathan Harrison Mar 2021

Bacterial Cyclic Diguanylate Signaling Networks Sense Temperature, Henrik Almblad, Trevor E. Randall, Fanny Liu, Katherine Leblanc, Ryan A. Groves, Weerayuth Kittichotirat, Geoffrey L. Winsor, Nicolas Fournier, Emily Au, Julie Groizeleau, Jacquelyn D. Rich, Yuefei Lou, Elise Granton, Laura K. Jennings, Larissa A. Singletary, Tara M.L. Winstone, Nathan M. Good, Roger E. Bumgarner, Michael F. Hynes, Manu Singh, Maria Silvina Stietz, Fiona S.L. Brinkman, Ayush Kumar, Ann Karen Cornelia Brassinga, Matthew R. Parsek, Boo Shan Tseng, Ian A. Lewis, Bryan G. Yipp, Justin L. Maccallum, Joe Jonathan Harrison

Life Sciences Faculty Research

Many bacteria use the second messenger cyclic diguanylate (c-di-GMP) to control motility, biofilm production and virulence. Here, we identify a thermosensory diguanylate cyclase (TdcA) that modulates temperature-dependent motility, biofilm development and virulence in the opportunistic pathogen Pseudomonas aeruginosa. TdcA synthesizes c-di-GMP with catalytic rates that increase more than a hundred-fold over a ten-degree Celsius change. Analyses using protein chimeras indicate that heat-sensing is mediated by a thermosensitive Per-Arnt-SIM (PAS) domain. TdcA homologs are widespread in sequence databases, and a distantly related, heterologously expressed homolog from the Betaproteobacteria order Gallionellales also displayed thermosensitive diguanylate cyclase activity. We propose, therefore, that thermotransduction …


Mfd Affects Global Transcription And The Physiology Of Stressed Bacillus Subtilis Cells, Holly Anne Martin, Anitha Sundararajan, Tatiana S. Ermi, Robert Heron, Jason Gonzales, Kaiden Lee, Diana Anguiano-Mendez, Faye Schilkey, Mario Pedraza-Reyes, Eduardo A. Robleto Jan 2021

Mfd Affects Global Transcription And The Physiology Of Stressed Bacillus Subtilis Cells, Holly Anne Martin, Anitha Sundararajan, Tatiana S. Ermi, Robert Heron, Jason Gonzales, Kaiden Lee, Diana Anguiano-Mendez, Faye Schilkey, Mario Pedraza-Reyes, Eduardo A. Robleto

Life Sciences Faculty Research

© Copyright © 2021 Martin, Sundararajan, Ermi, Heron, Gonzales, Lee, Anguiano-Mendez, Schilkey, Pedraza-Reyes and Robleto. For several decades, Mfd has been studied as the bacterial transcription-coupled repair factor. However, recent observations indicate that this factor influences cell functions beyond DNA repair. Our lab recently described a role for Mfd in disulfide stress that was independent of its function in nucleotide excision repair and base excision repair. Because reports showed that Mfd influenced transcription of single genes, we investigated the global differences in transcription in wild-type and mfd mutant growth-limited cells in the presence and absence of diamide. Surprisingly, we found …


Expanding Magnetic Organelle Biogenesis In The Domain Bacteria, Wei Lin, Wensi Zhang, Greig A. Paterson, Qiyun Zhu, Xiang Zhao, Rob Knight, Dennis A. Bazylinski, Andrew P. Roberts, Yongxin Pan Oct 2020

Expanding Magnetic Organelle Biogenesis In The Domain Bacteria, Wei Lin, Wensi Zhang, Greig A. Paterson, Qiyun Zhu, Xiang Zhao, Rob Knight, Dennis A. Bazylinski, Andrew P. Roberts, Yongxin Pan

Life Sciences Faculty Research

Background: The discovery of membrane-enclosed, metabolically functional organelles in Bacteria has transformed our understanding of the subcellular complexity of prokaryotic cells. Biomineralization of magnetic nanoparticles within magnetosomes by magnetotactic bacteria (MTB) is a fascinating example of prokaryotic organelles. Magnetosomes, as nano-sized magnetic sensors in MTB, facilitate cell navigation along the local geomagnetic field, a behaviour referred to as magnetotaxis or microbial magnetoreception. Recent discovery of novel MTB outside the traditionally recognized taxonomic lineages suggests that MTB diversity across the domain Bacteria are considerably underestimated, which limits understanding of the taxonomic distribution and evolutionary origin of magnetosome organelle biogenesis. Results: Here, …


Beneficial Endophytic Bacterial Populations Associated With Medicinal Plant Thymus Vulgaris Alleviate Salt Stress And Confer Resistance To Fusarium Oxysporum, Osama Aballa Abdelshafy Mohamad, Jin-Bao Ma, Yong-Hong Liu, Daoyuan Zhang, Shao Hua, Shirkant Bhute, Brian P. Hedlund, Wen-Jun Li, Li Li Feb 2020

Beneficial Endophytic Bacterial Populations Associated With Medicinal Plant Thymus Vulgaris Alleviate Salt Stress And Confer Resistance To Fusarium Oxysporum, Osama Aballa Abdelshafy Mohamad, Jin-Bao Ma, Yong-Hong Liu, Daoyuan Zhang, Shao Hua, Shirkant Bhute, Brian P. Hedlund, Wen-Jun Li, Li Li

Life Sciences Faculty Research

As a result of climate change, salinity has become a major abiotic stress that reduces plant growth and crop productivity worldwide. A variety of endophytic bacteria alleviate salt stress; however, their ecology and biotechnological potential has not been fully realized. To address this gap, a collection of 117 endophytic bacteria were isolated from wild populations of the herb Thymus vulgaris in Sheikh Zuweid and Rafah of North Sinai Province, Egypt, and identified based on their 16S rRNA gene sequences. The endophytes were highly diverse, including 17 genera and 30 species. The number of bacterial species obtained from root tissues was …


A Method For Improving The Accuracy And Efficiency Of Bacteriophage Genome Annotation, Alicia Salisbury, Philippos K. Tsourkas Jul 2019

A Method For Improving The Accuracy And Efficiency Of Bacteriophage Genome Annotation, Alicia Salisbury, Philippos K. Tsourkas

Life Sciences Faculty Research

Bacteriophages are the most numerous entities on Earth. The number of sequenced phage genomes is approximately 8000 and increasing rapidly. Sequencing of a genome is followed by annotation, where genes, start codons, and functions are putatively identified. The mainstays of phage genome annotation are auto-annotation programs such as Glimmer and GeneMark. Due to the relatively small size of phage genomes, many groups choose to manually curate auto-annotation results to increase accuracy. An additional benefit of manual curation of auto-annotated phage genomes is that the process is amenable to be performed by students, and has been shown to improve student recruitment …


The Anti-Sigma Factor Muca Of Pseudomonas Aeruginosa: Dramatic Differences Of A Muca22 Vs. A Δmuca Mutant In Anaerobic Acidified Nitrite Sensitivity Of Planktonic And Biofilm Bacteria In Vitro And During Chronic Murine Lung Infection, Warunya Panmanee, Shengchang Su, Michael J. Schurr, Gee W. Lau, Xiaoting Zhu, Zhaowei Ren, Cameron T. Mcdaniel, Long J. Lu, Dennis E. Ohman, Daniel A. Muruve, Ralph J. Panos, Hongwei D. Yu, Thomas B. Thompson, Boo Shan Tseng, Daniel J. Hassett Jun 2019

The Anti-Sigma Factor Muca Of Pseudomonas Aeruginosa: Dramatic Differences Of A Muca22 Vs. A Δmuca Mutant In Anaerobic Acidified Nitrite Sensitivity Of Planktonic And Biofilm Bacteria In Vitro And During Chronic Murine Lung Infection, Warunya Panmanee, Shengchang Su, Michael J. Schurr, Gee W. Lau, Xiaoting Zhu, Zhaowei Ren, Cameron T. Mcdaniel, Long J. Lu, Dennis E. Ohman, Daniel A. Muruve, Ralph J. Panos, Hongwei D. Yu, Thomas B. Thompson, Boo Shan Tseng, Daniel J. Hassett

Life Sciences Faculty Research

Mucoid mucA22 Pseudomonas aeruginosa (PA) is an opportunistic lung pathogen of cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) patients that is highly sensitive to acidified nitrite (A-NO2-). In this study, we first screened PA mutant strains for sensitivity or resistance to 20 mM A-NO2- under anaerobic conditions that represent the chronic stages of the aforementioned diseases. Mutants found to be sensitive to A-NO2- included PA0964 (pmpR, PQS biosynthesis), PA4455 (probable ABC transporter permease), katA (major catalase, KatA) and rhlR (quorum sensing regulator). In contrast, mutants lacking PA0450 (a putative phosphate transporter) and PA1505 (moaA2) were A-NO2- resistant. However, …


Uptake And Persistence Of Bacterial Magnetite Magnetosomes In A Mammalian Cell Line: Implications For Medical And Biotechnological Applications., Jefferson Cypriano, Jacques Werckmann, Gabriele Vargas, Adrana Lopes Dos Santos, Karen T. Silva, Pedro Leao, Fernando P. Almeida, Dennis A. Bazylinski, Marcos Farina, Ulysses Lins, Fernanda Abreu Apr 2019

Uptake And Persistence Of Bacterial Magnetite Magnetosomes In A Mammalian Cell Line: Implications For Medical And Biotechnological Applications., Jefferson Cypriano, Jacques Werckmann, Gabriele Vargas, Adrana Lopes Dos Santos, Karen T. Silva, Pedro Leao, Fernando P. Almeida, Dennis A. Bazylinski, Marcos Farina, Ulysses Lins, Fernanda Abreu

Life Sciences Faculty Research

Magnetotactic bacteria biomineralize intracellular magnetic nanocrystals surrounded by a lipid bilayer called magnetosomes. Due to their unique characteristics, magnetite magnetosomes are promising tools in Biomedicine. However, the uptake, persistence, and accumulation of magnetosomes within mammalian cells have not been well studied. Here, the endocytic pathway of magnetite magnetosomes and their effects on human cervix epithelial (HeLa) cells were studied by electron microscopy and high spatial resolution nano-analysis techniques. Transmission electron microscopy of HeLa cells after incubation with purified magnetosomes showed the presence of magnetic nanoparticles inside or outside endosomes within the cell, which suggests different modes of internalization, and that …


Genomic Analysis Of 48 Paenibacillus Larvae Bacteriophages, Casey Stamereilers, Christopher P. Fajardo, Jamison K. Walker, Katterinne N. Mendez, Eduardo Castro-Nallar, Julianne H. Grose, Sandra Hope, Philippos K. Tsourkas Jul 2018

Genomic Analysis Of 48 Paenibacillus Larvae Bacteriophages, Casey Stamereilers, Christopher P. Fajardo, Jamison K. Walker, Katterinne N. Mendez, Eduardo Castro-Nallar, Julianne H. Grose, Sandra Hope, Philippos K. Tsourkas

Life Sciences Faculty Research

The antibiotic-resistant bacterium Paenibacillus larvae is the causative agent of American foulbrood (AFB), currently the most destructive bacterial disease in honeybees. Phages that infect P. larvae were isolated as early as the 1950s, but it is only in recent years that P. larvae phage genomes have been sequenced and annotated. In this study we analyze the genomes of all 48 currently sequenced P. larvae phage genomes and classify them into four clusters and a singleton. The majority of P. larvaephage genomes are in the 38–45 kbp range and use the cohesive ends (cos) DNA-packaging strategy, while a minority have genomes …


Comparative Genomics Of 9 Novel Paenibacillus Larvae Bacteriophages, Casey Stamereilers, Lucy Leblanc, Diane Yost, Penny S. Amy, Philippos K. Tsourkas Aug 2016

Comparative Genomics Of 9 Novel Paenibacillus Larvae Bacteriophages, Casey Stamereilers, Lucy Leblanc, Diane Yost, Penny S. Amy, Philippos K. Tsourkas

Life Sciences Faculty Research

American Foulbrood Disease, caused by the bacterium Paenibacillus larvae, is one of the most destructive diseases of the honeybee, Apis mellifera. Our group recently published the sequences of 9 new phages with the ability to infect and lyse P. larvae. Here, we characterize the genomes of these P. larvae phages, compare them to each other and to other sequenced P. larvae phages, and putatively identify protein function. The phage genomes are 38–45 kb in size and contain 68–86 genes, most of which appear to be unique to P. larvae phages. We classify P. larvae phages into 2 main clusters and …


Microbial Endemism And Biogeography, Brian P. Hedlund, James T. Staley Jan 2004

Microbial Endemism And Biogeography, Brian P. Hedlund, James T. Staley

Life Sciences Faculty Research

The topic of microbial biogeography is almost 100 years old, however, when confronted with questions about the existence and extent of endemism in the microbial world, many microbiologists respond with opinions and theoretical arguments rather than examples of well-conducted studies. We begin this chapter with an overview of this debate as it applies to free-living prokayotes in part because there are relatively few good microbial biogeography studies. Furthermore, the arguments help to frame microbial biogeography in the larger context of biodiversity in that if endemism is common, then many more species exist.