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Transcription against an applied force

H Yin1, M D Wang, K Svoboda

  • 1Department of Biochemistry, Brandeis University, Waltham, MA 02254, USA.

Science (New York, N.Y.)
|December 8, 1995
PubMed
Summary

Researchers measured the force of a single Escherichia coli RNA polymerase molecule during transcription using optical trapping. This enzyme generates a force of 14 piconewtons, exceeding that of other molecular motors.

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Area of Science:

  • Molecular Biology
  • Biophysics
  • Biochemistry

Background:

  • Understanding the mechanical forces generated by molecular motors is crucial for elucidating cellular processes.
  • Escherichia coli RNA polymerase is a key enzyme responsible for transcription, a fundamental biological process.

Purpose of the Study:

  • To optically measure the force produced by a single molecule of Escherichia coli RNA polymerase during transcription.
  • To compare the forces generated by RNA polymerase with other known molecular motors.

Main Methods:

  • Utilized optical trapping interferometry to measure the force exerted by immobilized E. coli RNA polymerase.
  • Transcribed a DNA template attached to a polystyrene bead, measuring bead position to determine force.

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Main Results:

  • A single RNA polymerase molecule stalled reversibly at a mean applied force of 14 piconewtons at saturating nucleoside triphosphate concentrations.
  • The measured force is significantly greater than that produced by cytoskeletal motors like kinesin and myosin.
  • This force exceeds estimated mechanical loads opposing in vivo transcriptional elongation.

Conclusions:

  • The data suggest efficient conversion of free energy from RNA synthesis into mechanical work by RNA polymerase.
  • RNA polymerase generates substantial forces, highlighting its role as a powerful molecular motor in vivo.