Biomedical subjects

David Izhaky

Publications and source records attributed to David Izhaky.

3 recordsLinked to original sources

PubMed2006

Differential detection of dual traps improves the spatial resolution of optical tweezers.

The drive toward more sensitive single-molecule manipulation techniques has led to the recent development of optical tweezers capable of resolving the motions of biological systems at the subnanometer level, approaching the fundamental limit set by Brownian fluctuations. One successful approach has been the dual-trap optical tweezers, in which the system of study is held at both ends by microspheres in two separate optical traps. We present here a theoretical description of the Brownian limit on the spatial resolution of such systems and verify these predictions by direct measurement in a Brownian noise-limited dual-trap optical tweezers. We find that by detecting the positions of both trapped microspheres, correlations in their motions can be exploited to maximize the resolving power of the instrument. Remarkably, we show that the spatial resolution of dual optical traps with dual-trap detection is always superior to that of more traditional, single-trap designs, despite the added Brownian noise of the second trapped microsphere.

DNA↗

PubMed2004

Mechanical processes in biochemistry.

Mechanical processes are involved in nearly every facet of the cell cycle. Mechanical forces are generated in the cell during processes as diverse as chromosomal segregation, replication, transcription, translation, translocation of proteins across membranes, cell locomotion, and catalyzed protein and nucleic acid folding and unfolding, among others. Because force is a product of all these reactions, biochemists are beginning to directly apply external forces to these processes to alter the extent or even the fate of these reactions hoping to reveal their underlying molecular mechanisms. This review provides the conceptual framework to understand the role of mechanical force in biochemistry.

Animals↗

PubMed2002

Using mechanical force to probe the mechanism of pausing and arrest during continuous elongation by Escherichia coli RNA polymerase.

Escherichia coli RNA polymerase translocates along the DNA discontinuously during the elongation phase of transcription, spending proportionally more time at some template positions, known as pause and arrest sites, than at others. Current models of elongation suggest that the enzyme backtracks at these locations, but the dynamics are unresolved. Here, we study the role of lateral displacement in pausing and arrest by applying force to individually transcribing molecules. We find that an assisting mechanical force does not alter the translocation rate of the enzyme, but does reduce the efficiency of both pausing and arrest. Moreover, arrested molecules cannot be rescued by force, suggesting that arrest occurs by a bipartite mechanism: the enzyme backtracks along the DNA followed by a conformational change of the ternary complex (RNA polymerase, DNA and transcript), which cannot be reversed mechanically.

DNA-Directed RNA Polymerases↗