scholarly journals The specificity of ParR binding determines the compatibility of conjugative plasmids inClostridium perfringens

2018 ◽  
Author(s):  
Thomas D. Watts ◽  
Daouda A.K. Traore ◽  
Sarah C. Atkinson ◽  
Carmen Lao ◽  
Natalie Caltabiano ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Binding Sites ◽  
Selective Advantage ◽  
Dna Binding Proteins ◽  
Amino Acid Sequences ◽  
Host Bacterium ◽  
Replication Machinery ◽  
Rep Proteins ◽  
Resistance Plasmids ◽  
Encoding Genes

ABSTRACTPlasmids that encode the same replication machinery are generally unable to coexist in the same bacterial cell. However,Clostridium perfringensstrains often carry multiple conjugative toxin or antibiotic resistance plasmids that are closely related and encode similar Rep proteins. In many bacteria, plasmid partitioning upon cell division involves a ParMRC system and there are ~10 different ParMRC families inC. perfringens, with differences in amino acid sequences between each ParM family (15% − 54% identity). Since plasmids encoding genes belonging to the same ParMRC family are not observed in the same strain, these families appear to represent the basis for plasmid compatibility inC. perfringens. To understand this process, we examined the key recognition steps between ParR DNA-binding proteins and theirparCbinding sites. The ParR proteins bound to sequences within aparCsite from the same ParMRC family, but could not interact with aparCsite from a different ParMRC family. These data provide evidence that compatibility of the conjugative toxin plasmids ofC. perfringensis mediated by theirparMRC-like partitioning systems. This process provides a selective advantage by enabling the host bacterium to maintain separate plasmids that encode toxins that are specific for different host targets.

scholarly journals Exogenous Isolation of Antibiotic Resistance Plasmids from Piggery Manure Slurries Reveals a High Prevalence and Diversity of IncQ-Like Plasmids

2000 ◽  
Vol 66 (11) ◽  
pp. 4854-4862 ◽  
Author(s):  
Kornelia Smalla ◽  
Holger Heuer ◽  
Antje Götz ◽  
Dagmar Niemeyer ◽  
Ellen Krögerrecklenfort ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Ralstonia Eutropha ◽  
Blot Hybridization ◽  
Restriction Pattern ◽  
Pcr Analysis ◽  
Dot Blot ◽  
High Prevalence ◽  
Resistance Plasmids ◽  
Incq Plasmids ◽  
K 12

ABSTRACT Antibiotic resistance plasmids were exogenously isolated in biparental matings with piggery manure bacteria as plasmid donors inEscherichia coli CV601 and Pseudomonas putidaUWC1 recipients. Surprisingly, IncQ-like plasmids were detected by dot blot hybridization with an IncQ oriV probe in severalP. putida UWC1 transconjugants. The capture of IncQ-like plasmids in biparental matings indicates not only their high prevalence in manure slurries but also the presence of efficiently mobilizing plasmids. In order to elucidate unusual hybridization data (weak or no hybridization with IncQ repB or IncQ oriTprobes) four IncQ-like plasmids (pIE1107, pIE1115, pIE1120, and pIE1130), each representing a different EcoRV restriction pattern, were selected for a more thorough plasmid characterization after transfer into E. coli K-12 strain DH5α by transformation. The characterization of the IncQ-like plasmids revealed an astonishingly high diversity with regard to phenotypic and genotypic properties. Four different multiple antibiotic resistance patterns were found to be conferred by the IncQ-like plasmids. The plasmids could be mobilized by the RP4 derivative pTH10 into Acinetobactersp., Ralstonia eutropha, Agrobacterium tumefaciens, and P. putida, but they showed diverse patterns of stability under nonselective growth conditions in different host backgrounds. Incompatibility testing and PCR analysis clearly revealed at least two different types of IncQ-like plasmids. PCR amplification of total DNA extracted directly from different manure samples and other environments indicated the prevalence of both types of IncQ plasmids in manure, sewage, and farm soil. These findings suggest that IncQ plasmids play an important role in disseminating antibiotic resistance genes.

scholarly journals Identification, characterization and cloning of myr 1, a mammalian myosin-I.

1993 ◽  
Vol 120 (6) ◽  
pp. 1393-1403 ◽  
Author(s):  
C Ruppert ◽  
R Kroschewski ◽  
M Bähler
Keyword(s):  
Amino Acid ◽  
Light Chain ◽  
Brush Border ◽  
Binding Sites ◽  
Neuronal Development ◽  
Amino Acid Sequences ◽  
Dependent Manner ◽  
Tail Domain ◽  
Myosin I ◽  
Splice Forms

We have identified, characterized and cloned a novel mammalian myosin-I motor-molecule, called myr 1 (myosin-I from rat). Myr 1 exists in three alternative splice forms: myr 1a, myr 1b, and myr 1c. These splice forms differ in their numbers of putative calmodulin/light chain binding sites. Myr 1a-c were selectively released by ATP, bound in a nucleotide-dependent manner to F-actin and exhibited amino acid sequences characteristic of myosin-I motor domains. In addition to the motor domain, they contained a regulatory domain with up to six putative calmodulin/light chain binding sites and a tail domain. The tail domain exhibited 47% amino acid sequence identity to the brush border myosin-I tail domain, demonstrating that myr 1 is related to the only other mammalian myosin-I motor molecule that has been characterized so far. In contrast to brush border myosin-I which is expressed in mature enterocytes, myr 1 splice forms were differentially expressed in all tested tissues. Therefore, myr 1 is the first mammalian myosin-I motor molecule with a widespread tissue distribution in neonatal and adult tissues. The myr 1a splice form was preferentially expressed in neuronal tissues. Its expression was developmentally regulated during rat forebrain ontogeny and subcellular fractionation revealed an enrichment in purified growth cone particles, data consistent with a role for myr 1a in neuronal development.

Antibiotic resistance and OXA-type carbapenemases-encoding genes in airborne Acinetobacter baumannii isolated from burn wards

Burns ◽  
2014 ◽  
Vol 40 (2) ◽  
pp. 295-299 ◽  
Author(s):  
Jing Gao ◽  
Xiaonan Zhao ◽  
Ying Bao ◽  
Ruihua Ma ◽  
Yufa Zhou ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Acinetobacter Baumannii ◽  
Encoding Genes

scholarly journals Screening of a Library of Oligosaccharides Targeting Lectin LecB of Pseudomonas Aeruginosa and Synthesis of High Affinity Oligoglycoclusters

Molecules ◽  
2018 ◽  
Vol 23 (12) ◽  
pp. 3073 ◽  
Author(s):  
Lucie Dupin ◽  
Mathieu Noël ◽  
Silvère Bonnet ◽  
Albert Meyer ◽  
Thomas Géhin ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Binding Sites ◽  
Biofilm Formation ◽  
Selective Advantage ◽  
Host Cells ◽  
Host Defenses ◽  
Negative Bacterium ◽  
High Affinity ◽  
Structure Relationship ◽  
Relationship Of

The Gram negative bacterium Pseudomonas aeruginosa (PA) is an opportunistic bacterium that causes severe and chronic infection of immune-depressed patients. It has the ability to form a biofilm that gives a selective advantage to the bacteria with respect to antibiotherapy and host defenses. Herein, we have focused on the tetrameric soluble lectin which is involved in bacterium adherence to host cells, biofilm formation, and cytotoxicity. It binds to l-fucose, d-mannose and glycan exposing terminal fucose or mannose. Using a competitive assay on microarray, 156 oligosaccharides and polysaccharides issued from fermentation or from the biomass were screened toward their affinity to LecB. Next, the five best ligands (Lewisa, Lewisb, Lewisx, siayl-Lewisx and 3-fucosyllactose) were derivatized with a propargyl aglycon allowing the synthesis of 25 trivalent, 25 tetravalent and 5 monovalent constructions thanks to copper catalyzed azide alkyne cycloaddition. The 55 clusters were immobilized by DNA Directed immobilization leading to the fabrication of a glycocluster microarray. Their binding to LecB was studied. Multivalency improved the binding to LecB. The binding structure relationship of the clusters is mainly influenced by the carbohydrate residues. Molecular simulations indicated that the simultaneous contact of both binding sites of monomer A and D seems to be energetically possible.

scholarly journals Compensatory mutations improve general permissiveness to antibiotic resistance plasmids

2017 ◽  
Vol 1 (9) ◽  
pp. 1354-1363 ◽  
Author(s):  
Wesley Loftie-Eaton ◽  
Kelsie Bashford ◽  
Hannah Quinn ◽  
Kieran Dong ◽  
Jack Millstein ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Plasmids ◽  
Compensatory Mutations

scholarly journals Molecular Analysis of pDL10 from Acidianus ambivalens Reveals a Family of Related Plasmids from Extremely Thermophilic and Acidophilic Archaea

Genetics ◽  
1999 ◽  
Vol 152 (4) ◽  
pp. 1307-1314
Author(s):  
Arnulf Kletzin ◽  
Angelika Lieke ◽  
Tim Urich ◽  
Robert L Charlebois ◽  
Christoph W Sensen
Keyword(s):  
Amino Acid ◽  
Regulatory Protein ◽  
Amino Acid Sequences ◽  
Open Reading Frames ◽  
Mung Bean Nuclease ◽  
Stem Loop ◽  
Rolling Circle ◽  
Rep Protein ◽  
Homologous Protein ◽  
Rep Proteins

Abstract The 7598-bp plasmid pDL10 from the extremely thermophilic, acidophilic, and chemolithoautotrophic Archaeon Acidianus ambivalens was sequenced. It contains 10 open reading frames (ORFs) organized in five putative operons. The deduced amino acid sequence of the largest ORF (909 aa) showed similarity to bacterial Rep proteins known from phages and plasmids with rolling-circle (RC) replication. From the comparison of the amino acid sequences, a novel family of RC Rep proteins was defined. The pDL10 Rep protein shared 45-80% identical residues with homologous protein genes encoded by the Sulfolobus islandicus plasmids pRN1 and pRN2. Two DNA regions capable of forming extended stem-loop structures were also conserved in the three plasmids (48-69% sequence identity). In addition, a putative plasmid regulatory protein gene (plrA) was found, which was conserved among the three plasmids and the conjugative Sulfolobus plasmid pNOB8. A homolog of this gene was also found in the chromosome of S. solfataricus. Single-stranded DNA of both pDL10 strands was detected with a mung bean nuclease protection assay using PCR detection of protected fragments, giving additional evidence for an RC mechanism of replication.

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