The clamp-loader-helicase interaction in Bacillus: Leucine 381 is critical for pentamerisation and helicase-binding of the Bacillus tau protein.

A. Haroniti, R. Till, Margaret Caroline MacHin Smith, P. Soultanas

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Recently, we revealed the architecture of the clamp-loader-helicase (T-DnaB) complex in Bacillus by atomic force microscopy imaging and constructed a structural model, whereby a pentameric clamp-loader interacts with the hexameric helicase. Crucial to this model is the assumption that the clamp-loader forms a pentamer in the absence of other components of the clamp-loader complex such as deltadelta'. Here, we show that the Bacillus subtilis tau protein, even in the absence of deltadelta', interacts as a pentamer with the hexameric DnaB and that the L381 of tau is critical for the integrity of the tau oligomer and interaction with DnaB. The effects of the L381A mutation were confirmed by gel filtration, ultracentrifugation, circular dichroism, cross-linking studies, and genetic replacement of the dnaX gene with a mutant L381A dnaX gene in vivo. The L381A protein is able to support growth in vivo only when expressed in high quantities. Finally, despite the fact that a mutation at P465 has been reported to result in a thermosensitive gene in vivo, a P465L mutant protein interacts with DnaB in vitro suggesting that this defect is not a result of a defective T-DnaB, interaction.

Original languageEnglish
Pages (from-to)10955-10964
Number of pages9
JournalBiochemistry
Volume42
DOIs
Publication statusPublished - 2003

Keywords

  • POLYMERASE-III HOLOENZYME
  • COLI REPLICATION PROTEINS
  • UNIQUE C-TERMINUS
  • DNA-REPLICATION
  • DISTINCT DOMAINS
  • SLIDING CLAMPS
  • READING FRAME
  • DELTA-DELTA'
  • SUBUNIT
  • COMPLEX

Cite this

The clamp-loader-helicase interaction in Bacillus: Leucine 381 is critical for pentamerisation and helicase-binding of the Bacillus tau protein. / Haroniti, A.; Till, R.; Smith, Margaret Caroline MacHin; Soultanas, P.

In: Biochemistry, Vol. 42, 2003, p. 10955-10964.

Research output: Contribution to journalArticle

Haroniti, A. ; Till, R. ; Smith, Margaret Caroline MacHin ; Soultanas, P. / The clamp-loader-helicase interaction in Bacillus: Leucine 381 is critical for pentamerisation and helicase-binding of the Bacillus tau protein. In: Biochemistry. 2003 ; Vol. 42. pp. 10955-10964.
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