ABSTRACT N , -Dimethylformamide (DMF), a toxic solvent used in the chemical industry, is frequently present industrial wastes. Plasmid pAMI2 (18.6 kb) of Paracoccus aminophilus JCM 7686 carries genetic information which crucial for methylotrophic growth this bacterium, using DMF as sole source carbon and energy. Besides conserved backbone related to pAgK84 Agrobacterium radiobacter K84, three-gene cluster coding protein DmfR, has sequence similarities members LuxR family transcription...

Plasmid conjugative transfer systems comprise type IV secretion (T4SS) coupled to DNA processing and replication. The T4SSs are divided into two phylogenetic subfamilies, namely, IVA IVB, or on the basis of phylogeny VirB4 ATPase, eight groups. conjugation system IncM group plasmid pCTX-M3, from Citrobacter freundii, is classified in IVB subfamily MPFI group, as IncI1 plasmids. Although majority genes plasmids display conserved synteny, there several differences. Here, we present a deletion...

High segregational stability of the streptococcal plasmid pSM19035 is achieved by concerted action systems involved in copy number control, multimer resolution, and postsegregational killing. In this study, we demonstrate role two genes, delta omega, stabilization a partition mechanism. We show that these genes can stabilize native replicon as well other theta- sigma-type plasmids Bacillus subtilis. contrast to known systems, case are transcribed separately; however, they coregulated product...

Low copy number plasmids cannot rely on the random segregation during bacterial cell division. To be stably maintained in population they evolved two types of mechanisms (i) partition systems (PAR) that actively separate replicated plasmid molecules to daughter cells and (ii) toxin-andidote (TA) act after division kill plasmid-less cells. Our knowledge has been based mainly analysis from Gram-negative bacteria. Now, numerous Gram-positive bacteria have also characterized make significant...

Faithful replication and repair of DNA lesions ensure genome maintenance. During in eukaryotic cells, is unwound by the CMG helicase complex, which composed three major components: Cdc45 protein, Mcm2-7, GINS complex. The complex with polymerase epsilon (CMG-E) participates establishment progression replisome. Impaired functioning CMG-E was shown to induce genomic instability promote development various diseases. Therefore, components play important roles as caretakers genome. In...

Abstract Cdc45 is a non-catalytic subunit of the CMG helicase complex and recruited to autonomously replicating sequence at onset DNA replication. protein required for initiation replication process as well nascent strand synthesis. It interacts with Mcm2 Psf1 elements helicase, Sld3, factor, Pol2, catalytic polymerase epsilon (Pol ε). In this study, we analyzed effects amino acid substitutions in regions involved interaction Mcm2-7 (Cdc45-1), (Cdc45-26), Sld3 (Cdc45-25, Cdc45-35). We found...

To preserve genome integrity, the S-phase checkpoint senses damaged DNA or nucleotide depletion and when necessary, arrests replication progression delays cell division. Previous studies, based on two pol2 mutants have suggested involvement of polymerase epsilon (Pol ε) in sensing accuracy Saccharomyces cerevisiae. Here we studied Pol ε proper replication, using a mutant DPB2, gene coding for non-catalytic subunit ε. Under genotoxic conditions, dpb2-103 cells progress through S phase faster...

The generally accepted model assumes that leading strand synthesis is performed by Pol ε, while lagging-strand catalyzed δ. ε has been shown to target the interacting with CMG helicase [Cdc45 Mcm2-7 GINS(Psf1-3, Sld5)]. Proper functioning of CMG-Pol ɛ, helicase-polymerase complex essential for its progression and fidelity DNA replication. Dpb2p, non-catalytic subunit plays a key role in maintaining correct architecture replisome acting as link between complex. Using temperature-sensitive...

Active support is needed for low copy-number plasmids to be stably maintained in bacterial cells. The mechanisms that fulfill this role are (i) partition systems (PAR) acting separate plasmid molecules daughter cells and (ii) toxin-andidote (TA) (post-segregational killing-PSK) which arrest cell growth until the reaches correct or kill have not inherited plasmid. Our knowledge of toxin-antidote comes mainly from studies on Gram-negative bacteria. However, some addiction Gram-positive...

DNA replication is performed by replisome proteins, which are highly conserved from yeast to humans. The CMG [Cdc45-Mcm2-7-GINS(Psf1-3, Sld5)] helicase unwinds the double helix separate leading and lagging strands, replicated specialized polymerases epsilon (Pol ε) delta δ), respectively. This division of labor was confirmed both genetic analyses in vitro studies. Exceptions this rule were described mainly cells with impaired catalytic polymerase ε subunit. central role recruitment...

Formation of the segrosome, a nucleoprotein complex crucial for proper functioning plasmid partition systems, involves interactions between specific proteins (ParA-like and ParB-like), ATP DNA sequences (the centromeric sites). Although systems have been studied many years, details segrosome formation are not yet clear. Organization pSM19035-encoded system is unique; in contrast with other known par here, δ ω genes do constitute an operon. Moreover, Omega [a ParB-like protein which has...

The CMG complex (Cdc45, Mcm2–7, GINS (Psf1, 2, 3, and Sld5)) is crucial for both DNA replication initiation fork progression. helicase interaction with the leading strand polymerase epsilon (Pol ε) essential preferential loading of Pol ε onto strand, stimulation polymerase, modulation activity. Here, we analyze consequences impaired between in Saccharomyces cerevisiae cells psf1-100 mutation. This significantly affects activity measured vitro, while vivo, mutation reduces fidelity by...

The conjugation of donor and recipient bacterial cells resulting in conjugative transfer mobilizable plasmids is the preferred method enabling introduction DNA into strains for which other methods are difficult to establish (e.g., clinical strains). We have constructed E. coli carrying system IncM plasmid pCTX-M3 integrated chromosome. To increase mobilization efficiency up 1,000-fold, two putative regulators this system, orf35 orf36 , were disabled. broaden repertoire tools Gram-negative...

Partition systems contribute to stable plasmid inheritance in bacteria through the active separation of DNA molecules daughter cells, and centromeric sequence located either upstream or downstream canonical partition operons plays an important role this process. A specific DNA-binding protein binds interacts with motor NTPase form a nucleoprotein complex. The inc18-family pSM19035 is partitioned by products δ ω genes, encoding Walker-type ATPase protein. As two genes are transcribed...

DNA replication in Saccharomyces cerevisiae and other eukaryotes is performed mainly by polymerase epsilon (Pol ε) on the leading strand delta δ) lagging strand. Using a mutant form of enables tracking its signature replicated DNA. Here, we used pol2-M644G allele encoding catalytic subunit Pol ε to analyse contribution yeast with psf1-1 an essential gene GINS complex. involved recruitment ε, major replicase. Thus, defective functioning can affect involvement replication. Together Cdc45...