Biomedical subjects

Eric Becker

Publications and source records attributed to Eric Becker.

5 recordsLinked to original sources

PubMed2006

DNA segregation by the bacterial actin AlfA during Bacillus subtilis growth and development.

We here identify a protein (AlfA; actin like filament) that defines a new family of actins that are only distantly related to MreB and ParM. AlfA is required for segregation of Bacillus subtilis plasmid pBET131 (a mini pLS32-derivative) during growth and sporulation. A 3-kb DNA fragment encoding alfA and a downstream gene (alfB) is necessary and sufficient for plasmid stability. AlfA-GFP assembles dynamic cytoskeletal filaments that rapidly turn over (t(1/2)< approximately 45 s) in fluorescence recovery after photobleaching experiments. A point mutation (alfA D168A) that completely inhibits AlfA subunit exchange in vivo is strongly defective for plasmid segregation, demonstrating that dynamic polymerization of AlfA is necessary for function. During sporulation, plasmid segregation occurs before septation and independently of the DNA translocase SpoIIIE and the chromosomal Par proteins Soj and Spo0J. The absence of the RacA chromosome anchoring protein reduces the efficiency of plasmid segregation (by about two-fold), suggesting that it might contribute to anchoring the plasmid at the pole during sporulation. Our results suggest that the dynamic polymerization of AlfA mediates plasmid separation during both growth and sporulation.

Actin Cytoskeleton↗

PubMed2006

Flk prevents premature secretion of the anti-sigma factor FlgM into the periplasm.

The flk locus of Salmonella typhimurium was identified as a regulator of flagellar gene expression in strains defective in P- and l-ring formation. Flk acts as a regulator of flagellar gene expression by modulating the protein levels of the anti-sigma28 factor FlgM. Evidence is presented which suggests that Flk is a cytoplasmic-facing protein anchored to the inner membrane by a single, C-terminal transmembrane-spanning domain (TMS). The specific amino acid sequence of the TMS is not essential for Flk activity, but membrane anchoring is essential. Membrane fractionation and visualization of protein fusions of green fluorescent protein derivatives to Flk suggested that the Flk protein is present in the membrane as punctate spots in number that are much greater than the number of flagellar basal structures. The turnover of the anti-sigma28 factor FlgM was increased in flk mutant strains. Using FlgM-beta-lactamase fusions we show the increased turnover of FlgM in flk null mutations is due to FlgM secretion into the periplasm where it is degraded. Our data suggest that Flk inhibits FlgM secretion by acting as a braking system for the flagellar-associated type III secretion system. A model is presented to explain a role for Flk in flagellar assembly and gene regulatory processes.

Bacterial Proteins↗

PubMed2005

Elements in the co-evolution of relaxases and their origins of transfer.

The central elements in the conjugative mobilization of most plasmids are the relaxase and its cognate origin of transfer (oriT). The relaxase of the plasmid R1162, together with its oriT, belong to a large and widely distributed family of related relaxase/oriT pairs. Several of the properties of these elements are considered for R1162 and for other members of this family with a view to understanding how systems for mobilization might have evolved.

Conjugation, Genetic↗

PubMed2003

Chromosome segregation in Eubacteria.

It is now clear that bacterial chromosomes rapidly separate in a manner independent of cell elongation, suggesting the existence of a mitotic apparatus in bacteria. Recent studies of bacterial cells reveal filamentous structures similar to the eukaryotic cytoskeleton, proteins that mediate polar chromosome anchoring during Bacillus subtilis sporulation, and SMC interacting proteins that are involved in chromosome condensation. A picture is thereby developing of how bacterial chromosomes are organized within the cell, how they are separated following duplication, and how these processes are coordinated with the cell cycle.

Bacteria↗

PubMed2002

Conjugative DNA synthesis: R1162 and the question of rolling-circle replication.

Strand-replacement synthesis during conjugative mating has been characterized by introducing into donor cells R1162 plasmid DNA containing a base-pair mismatch. Conjugative synthesis in donors occurs in the absence of vegetative plasmid replication, but with a lag between rounds of transfer, and with most strands being initiated at the normal site within the replicative origin. These characteristics argue against the idea that multiple plasmid copies are generated for successive rounds of transfer by rolling-circle replication. However, the R1162 relaxase protein can process molecules containing multiple transfer origins in the manner expected for the conversion of single-strand multimers, generated by rolling-circle replication, to unit-length molecules. This capability appears to be the result of a secondary cleavage reaction carried out by the protein. The possibility is raised that the processing of molecules with more than one origin of transfer might be a repair mechanism directed against adventitious DNA synthesis during transfer.

Base Pair Mismatch↗