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

Kohji Yamamoto, Syed Muniruzzaman, Malini Rajagopalan, Murty V V S Madiraju

Research output: Contribution to journalArticle

21 Citations (Scopus)

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 nucleotides and membrane phospholipids, and the latter helps to ensure that only the ATP form of the DnaA protein interacts continuously with oriC.

Original languageEnglish
Pages (from-to)305-311
Number of pages7
JournalBiochemical Journal
Volume363
Issue number2
DOIs
Publication statusPublished - Apr 15 2002
Externally publishedYes

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Adenine Nucleotides
Replication Origin
Mycobacterium tuberculosis
Phospholipids
Adenosine Diphosphate
Adenosine Triphosphate
Modulation
Proteins
Membranes
Phosphatidylglycerols
Cardiolipins
Micelles
Phosphatidylinositols
DNA Replication
Adenosine Triphosphatases
Hydrolysis
Nucleotides
DNA

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Modulation of mycobacterium tuberculosis DnaA protein-adenine-nucleotide interactions by acidic phospholipids. / Yamamoto, Kohji; Muniruzzaman, Syed; Rajagopalan, Malini; Madiraju, Murty V V S.

In: Biochemical Journal, Vol. 363, No. 2, 15.04.2002, p. 305-311.

Research output: Contribution to journalArticle

Yamamoto, Kohji ; Muniruzzaman, Syed ; Rajagopalan, Malini ; Madiraju, Murty V V S. / Modulation of mycobacterium tuberculosis DnaA protein-adenine-nucleotide interactions by acidic phospholipids. In: Biochemical Journal. 2002 ; Vol. 363, No. 2. pp. 305-311.
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