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Microbiology 153 (2007), 4050-4060; DOI  10.1099/mic.0.2007/011619-0
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Microbiology 153 (2007), 4050-4060; DOI  10.1099/mic.0.2007/011619-0
© 2007 Society for General Microbiology

Characterization of the mycobacterial chromosome segregation protein ParB and identification of its target in Mycobacterium smegmatis

Dagmara Jakimowicz1,2, Anna Brzostek3, Anna Rumijowska-Galewicz3, Paulina Zydek1, Alicja Dolzblasz1, Aleksandra Smulczyk-Krawczyszyn1, Tomasz Zimniak1, Lukasz Wojtasz1, Anna Zawilak-Pawlik1, Agnieszka Kois1, Jaroslaw Dziadek3 and Jolanta Zakrzewska-Czerwinska1,2

1 Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, ul. Weigla 12, 53-114 Wroclaw, Poland
2 Faculty of Biotechnology, University of Wroclaw, ul. Tamka 2, 50-137 Wroclaw, Poland
3 Medical Biology Center, Polish Academy of Sciences, Lodowa 106, 93-232 Lódz, Poland

Correspondence
Jolanta Zakrzewska-Czerwinska
zakrzew{at}iitd.pan.wroc.pl

Bacterial chromosomes (though not Escherichia coli and some other {gamma}-proteobacterial chromosomes) contain parS sequences and parAB genes encoding partitioning proteins, i.e. ParA (ATPase) and ParB (DNA-binding proteins) that are components of the segregation machinery. Here, mycobacterial parABS elements were characterized for the first time. parAB genes are not essential in Mycobacterium smegmatis; however, elimination or overexpression of ParB protein causes growth inhibition. Deletion of parB also leads to a rather severe chromosome segregation defect: up to 10 % of the cells were anucleate. Mycobacterial ParB protein uses three oriC-proximal parS sequences as targets to organize the origin region into a compact nucleoprotein complex. Formation of such a complex involves ParB–ParB interactions and is assisted by ParA protein.

Abbreviations: EMSA, electrophoretic mobility shift assay; GST, glutathione S-transferase; Ms, M. smegmatis; Mt, M. tuberculosis

Three supplementary figures showing the construction of the M. smegmatis {Delta}parB mutant, purified mycobacterial ParAB proteins and protein sequence alignment of homologous ParB proteins, and a supplementary table listing the oligonucleotides used in this study are available with the online version of this paper.




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A. A. Bartosik, J. Mierzejewska, C. M. Thomas, and G. Jagura-Burdzy
ParB deficiency in Pseudomonas aeruginosa destabilizes the partner protein ParA and affects a variety of physiological parameters
Microbiology, April 1, 2009; 155(4): 1080 - 1092.
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