Related Experiment Videos

Characterization of the mIHF gene of Mycobacterium smegmatis

M L Pedulla1, G F Hatfull

  • 1Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.

Journal of Bacteriology
|October 10, 1998
PubMed

Insights

Mycobacteriophage L5 integration depends on mycobacterial integration host factor (mIHF). This heat-stable protein is crucial for Mycobacterium smegmatis viability and most abundant before stationary phase.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Virology

Background:

  • Mycobacteriophage L5 integration into the host genome requires specific host factors.
  • The precise molecular mechanisms and host factors involved in mycobacteriophage integration are not fully elucidated.

Purpose of the Study:

  • To identify and characterize the host factor essential for mycobacteriophage L5 integration in vitro.
  • To investigate the properties and abundance of this essential host factor in Mycobacterium smegmatis.

Main Methods:

  • In vitro integration assays using mycobacteriophage L5 and Mycobacterium smegmatis extracts.
  • Protein purification and characterization of the identified integration host factor (mIHF).
  • Quantification of mIHF levels during different growth phases of Mycobacterium smegmatis.

Main Results:

  • Mycobacterial integration host factor (mIHF), a 105-residue heat-stable polypeptide, was identified as essential for in vitro integration of mycobacteriophage L5.
  • mIHF shows no obvious structural or sequence similarity to known HU or other small DNA-binding proteins.
  • mIHF abundance is highest just prior to Mycobacterium smegmatis entering stationary phase.

Conclusions:

  • Mycobacterial integration host factor (mIHF) is a novel and essential protein for mycobacteriophage L5 integration.
  • mIHF plays a critical role in the lifecycle of mycobacteriophage L5 and is vital for the viability of Mycobacterium smegmatis.

Related Concept Videos