Author : Natalie Anne Dye
Publisher : Stanford University
ISBN 13 :
Total Pages : 286 pages
Book Rating : 4.F/5 ( download)
Book Synopsis The Assembly and Interactions of MreB in the Maintenance of Cell Shape in Caulobacter Crescentus by : Natalie Anne Dye
Download or read book The Assembly and Interactions of MreB in the Maintenance of Cell Shape in Caulobacter Crescentus written by Natalie Anne Dye and published by Stanford University. This book was released on 2010 with total page 286 pages. Available in PDF, EPUB and Kindle. Book excerpt: This work focuses on the mechanism by which MreB contributes to the maintenance of cell shape in the gram-negative alpha-proteobacterium Caulobacter crescentus. The gene mreB encodes a protein that resembles actin, a eukaryotic cytoskeletal protein. Previously, it was shown that mreB is required to maintain a rod-like shape and localizes to a helical pattern near the cytoplasmic membrane. Here, we show that MreB is associated with regions of active growth in Caulobacter, as mutant strains that mislocalize MreB to the cell poles direct new growth at or near the poles. We present evidence to suggest that MreB contributes to the determination of proper length, width, and curvature through partially distinct mechanisms. The determination of proper width involves the essential proteins MreC and Pbp2, which are encoded in the mreB operon. While MreB and MreC are both required to position the cell wall transpeptidase Pbp2 along the lateral sidewalls and away from midcell, the two do not colocalize and each can maintain its localization in the absence of the other. When MreB is mislocalized to the poles, MreC and Pbp2 do not follow. These data argue against the idea that MreB provides a scaffold-like structure to localize enzymes that directly modify the cell wall. The determination of proper curvature, involves the intermediate filament-like protein, Crescentin. We identify a putative binding site on MreB for Crescentin or other curvature-mediating factors. We also show that the extent to which the subcellular localization of MreB changes over the cell cycle is correlated with cell size, indicating that MreB is involved in the coordination between elongation and division. In addition, we show that in vitro purified MreB spontaneously forms very stable polymers in the presence or absence of nucleotide. These polymers are globular or amorphous and only filamentous when placed on a highly positively charged surface of Poly-L-lysine. These in vitro data suggest that MreB is likely to be regulated at the disassembly step in the cell and that the cellular environment may influence the structure of MreB polymers. Lastly, we present biochemical evidence to support the existence of a disassembly factor in cytoplasmic Caulobacter extract. Together our data suggest that the maintenance of the crescent-rod cell shape in Caulobacter is the result of a complicated balance between MreB's dynamic subcellular localization, polymeric structure, and communication with cellular components.
