Modulation of Mycobacterium tuberculosis DnaA protein-adenine-nucleotide interactions by acidic phospholipids

The Biochemical Journal
Kohji YamamotoMurty Madiraju

Abstract

The biochemical aspects of the initiation of DNA replication in Mycobacterium tuberculosis are unknown. To understand this process, we overproduced, purified and characterized the recombinant M. tuberculosis DnaA protein. The M. tuberculosis DnaA protein binds the origin of replication (oriC), ATP and ADP, and exhibited weak ATPase activity. ADP, after hydrolysis of ATP, remained strongly associated with DnaA and the exchange of ATP for bound ADP was weak. Vesicles prepared from acidic phospholipids, such as phosphatidylinositol, cardiolipin and phosphatidylglycerol, promoted dissociation of both ADP and ATP, whereas the neutral phospholipid phosphatidylethanolamine did not. The phospholipid-mediated dissociation of ATP was decreased in the presence of the M. tuberculosis oriC, whereas dissociation of ADP was stimulated in the presence of oriC. Acidic phospholipids in micelles, however, were not efficient in dissociating bound nucleotides from DnaA. Together, these results suggest that both polar head groups and membrane bilayer structure play an important role in M. tuberculosis DnaA-adenine-nucleotide interactions. We suggest that initiation of M. tuberculosis oriC involves intimate interactions between DnaA, adenine nucleoti...Continue Reading

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Citations

Jul 24, 2008·Proceedings of the National Academy of Sciences of the United States of America·Stéphane DuigouMatthew K Waldor
Feb 26, 2008·Journal of Biochemistry·Kohji YamamotoMurty V V S Madiraju
Oct 22, 2010·Doklady Biological Sciences : Proceedings of the Academy of Sciences of the USSR, Biological Sciences Sections·S N AndreevskayaV I Shvets
Dec 7, 2011·Tuberculosis·Erin MaloneyMurty Madiraju
Jul 25, 2015·Molecular Microbiology·Gorla PurushothamMurty V Madiraju
Mar 24, 2006·Molecular Microbiology·Murty V V S MadirajuMalini Rajagopalan
Mar 7, 2006·Protein Expression and Purification·Anna M Zawilak-PawlikJolanta Zakrzewska-Czerwinska
Apr 27, 2007·FEMS Microbiology Reviews·Jolanta Zakrzewska-CzerwińskaWalter Messer
Jan 1, 2012·Life·Vic Norris, Patrick Amar

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