Dynamic Instability in a DNA-Segregating Prokaryotic Actin Homolog

Dynamic instability—the switching of a two-state polymer between phases of steady elongation and rapid shortening—is essential to the cellular function of eukaryotic microtubules, especially during chromosome segregation. Since the discovery of dynamic instability 20 years ago, no other biological polymer has been found to exhibit this behavior. Using total internal reflection fluorescence microscopy and fluorescence resonance energy transfer, we observe that the prokaryotic actin homolog ParM, whose assembly is required for the segregation of large, low–copy number plasmids, displays both dynamic instability and symmetrical, bidirectional polymerization. The dynamic instability of ParM is regulated by adenosine triphosphate (ATP) hydrolysis, and filaments are stabilized by a cap of ATP-bound monomers. ParM is not related to tubulin, so its dynamic instability must have arisen by convergent evolution driven by a set of common constraints on polymer-based segregation of DNA.

[1]  Elmer S. West From the U. S. A. , 1965 .

[2]  Peter Roepstorff,et al.  Bacterial mitosis: ParM of plasmid R1 moves plasmid DNA by an actin-like insertional polymerization mechanism. , 2003, Molecular cell.

[3]  T. Mitchison,et al.  Rate-limiting guanosine 5'-triphosphate hydrolysis during nucleotide turnover by FtsZ, a prokaryotic tubulin homologue involved in bacterial cell division. , 2004, Biochemistry.

[4]  P. Dancker,et al.  Influence of phalloidin on both the nucleation and the elongation phase of actin polymerization. , 1987, Biochimica et biophysica acta.

[5]  W. Baumeister,et al.  Supporting online material Materials and methods , 2002 .

[6]  T. Pollard,et al.  Hydrolysis of ATP by polymerized actin depends on the bound divalent cation but not profilin. , 2002, Biochemistry.

[7]  R. B. Jensen,et al.  Partitioning of plasmid R1. The ParM protein exhibits ATPase activity and interacts with the centromere-like ParR-parC complex. , 1997, Journal of molecular biology.

[8]  M. Kirschner,et al.  The minimum GTP cap required to stabilize microtubules , 1994, Current Biology.

[9]  F. Young Biochemistry , 1955, The Indian Medical Gazette.

[10]  Zemer Gitai,et al.  An actin-like gene can determine cell polarity in bacteria. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[11]  S. Molin,et al.  Partitioning of plasmid R1. Structural and functional analysis of the parA locus. , 1986, Journal of molecular biology.

[12]  S. Molin,et al.  Stable inheritance of plasmid R1 requires two different loci , 1985, Journal of bacteriology.

[13]  T. Pollard,et al.  Interactions of Acanthamoeba profilin with actin and nucleotides bound to actin. , 1998, Biochemistry.

[14]  P. Graumann,et al.  Actin-like Proteins MreB and Mbl from Bacillus subtilis Are Required for Bipolar Positioning of Replication Origins , 2003, Current Biology.

[15]  Charles Boone,et al.  Mechanism of formin-induced nucleation of actin filaments. , 2003, Biochemistry.

[16]  Jan Löwe,et al.  F‐actin‐like filaments formed by plasmid segregation protein ParM , 2002, The EMBO journal.

[17]  T. Kruse,et al.  Dysfunctional MreB inhibits chromosome segregation in Escherichia coli , 2003, The EMBO journal.

[18]  K. Nordström,et al.  Mutations in R Factors of Escherichia coli Causing an Increased Number of R-Factor Copies per Chromosome , 1972, Journal of bacteriology.

[19]  R. Mullins,et al.  Cellular control of actin nucleation. , 2002, Annual review of cell and developmental biology.

[20]  M. Carlier,et al.  Probing the mechanism of ATP hydrolysis on F-actin using vanadate and the structural analogs of phosphate BeF-3 and A1F-4. , 1988, The Journal of biological chemistry.

[21]  S. Almo,et al.  The structure of nonvertebrate actin: Implications for the ATP hydrolytic mechanism , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[22]  E. Nishida,et al.  Kinetic analysis of actin polymerization. , 1983, Journal of biochemistry.

[23]  T. Mitchison,et al.  Mitosis: a history of division , 2001, Nature Cell Biology.

[24]  M. Kirschner,et al.  Dynamic instability of microtubule growth , 1984, Nature.

[25]  S. Zigmond Formin-induced nucleation of actin filaments. , 2004, Current opinion in cell biology.

[26]  宁北芳,et al.  疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A , 2005 .

[27]  F. Oosawa,et al.  A theory of linear and helical aggregations of macromolecules. , 1962, Journal of molecular biology.

[28]  R. B. Jensen,et al.  Prokaryotic DNA segregation by an actin‐like filament , 2002, The EMBO journal.

[29]  E. Mandelkow,et al.  Dynamics of the microtubule oscillator: role of nucleotides and tubulin‐MAP interactions. , 1988, The EMBO journal.

[30]  K. Gerdes,et al.  Partitioning of plasmid R1. Ten direct repeats flanking the parA promoter constitute a centromere-like partition site parC, that expresses incompatibility. , 1994, Journal of molecular biology.

[31]  R. B. Jensen,et al.  Mechanism of DNA segregation in prokaryotes: ParM partitioning protein of plasmid R1 co‐localizes with its replicon during the cell cycle , 1999, The EMBO journal.

[32]  D. Panda,et al.  Determination of the size and chemical nature of the stabilizing "cap" at microtubule ends using modulators of polymerization dynamics. , 2002, Biochemistry.

[33]  T. L. Hill,et al.  Synchronous oscillations in microtubule polymerization. , 1987, Proceedings of the National Academy of Sciences of the United States of America.