Abstract
Condensins in bacteria are one of the most important factors involved in the organization of long threads of DNA into compact chromosomes. The organization of DNA by condensins is vital to many DNA transactions including DNA repair and chromosome segregation. Although some of the activities of condensins are well studied, the mechanism of the overall process executed by condensins, DNA compaction, remains unclear. Here, we describe some of the methods used routinely in our laboratory to understand the mechanism of DNA compaction by Escherichia coli condensin MukB.
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References
Uhlmann F (2016) SMC complexes: from DNA to chromosomes. Nat Rev Mol Cell Biol 17:399–412
Hirano T (2016) Condensin-based chromosome organization from bacteria to vertebrates. Cell 164:847–857
Wells JN, Gligoris TG, Nasmyth KA, Marsh JA (2017) Evolution of condensin and cohesin complexes driven by replacement of Kite by Hawk proteins. Curr Biol 27:R17–R18
Niki H, Jaffe A, Imamura R, Ogura T, Hiraga S (1991) The new gene mukB codes for a 177 kd protein with coiled-coil domains involved in chromosome partitioning of E. coli. EMBO J 10:183–193
Yamanaka K, Ogura T, Niki H, Hiraga S (1996) Identification of two new genes, mukE and mukF, involved in chromosome partitioning in Escherichia coli. Mol Gen Genet 250:241–251
Petrushenko ZM, Cui Y, She W, Rybenkov VV (2010) Mechanics of DNA bridging by bacterial condensin MukBEF in vitro and in singulo. EMBO J 29:1126–1135
Petrushenko ZM, Lai CH, Rybenkov VV (2006) Antagonistic interactions of kleisins and DNA with bacterial Condensin MukB. J Biol Chem 281:34208–34217
Bahng S, Hayama R, Marians KJ (2016) MukB-mediated catenation of DNA is ATP and MukEF independent. J Biol Chem 291:23999–24008
Zawadzka K, Zawadzki P, Baker R, Rajasekar KV, Wagner F, Sherratt DJ, Arciszewska LK (2018) MukB ATPases are regulated independently by the N- and C-terminal domains of MukF kleisin. elife 7:e31522
Kumar R, Grosbart M, Nurse P, Bahng S, Wyman CL, Marians KJ (2017) The bacterial condensin MukB compacts DNA by sequestering supercoils and stabilizing topologically isolated loops. J Biol Chem 292:16904–16920
Hayama R, Marians KJ (2010) Physical and functional interaction between the condensin MukB and the decatenase topoisomerase IV in Escherichia coli. Proc Natl Acad Sci U S A 107:18826–18831
Mizuuchi K, Mizuuchi M, O’Dea MH, Gellert M (1984) Cloning and simplified purification of Escherichia coli DNA gyrase A and B proteins. J Biol Chem 259:9199–9201
Hiasa H, DiGate RJ, Marians KJ (1994) Decatenating activity of Escherichia coli DNA gyrase and topoisomerases I and III during oriC and pBR322 DNA replication in vitro. J Biol Chem 269:2093–2099
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Kumar, R., Bahng, S., Marians, K.J. (2019). Dissecting DNA Compaction by the Bacterial Condensin MukB. In: Badrinarayanan, A. (eds) SMC Complexes. Methods in Molecular Biology, vol 2004. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9520-2_13
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DOI: https://doi.org/10.1007/978-1-4939-9520-2_13
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