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Jordanka Zlatanova

Publications and source records attributed to Jordanka Zlatanova.

6 recordsLinked to original sources
PubMed2003

Chromatin fibers, one-at-a-time.

Eukaryotic DNA is presented to the enzymatic machineries that use DNA as a template in the form of chromatin fibers. At the first level of organization, DNA is wrapped around histone octamers to form nucleosomal particles that are connected with stretches of linker DNA; this beads-on-a-string structure folds further to reach a very compact state in the nucleus. Chromatin structure is in constant flux, changing dynamically to accommodate the needs of the cell to replicate, transcribe, and repair the DNA, and to regulate all these processes in time and space. The more conventional biochemical and biophysical techniques used to study chromatin structure and dynamics have been recently complemented by an array of single-molecule approaches, in which chromatin fibers are investigated one-at-a-time. Here we describe single-molecule efforts to see nucleosomes, touch them, put them together, and then take them apart, one-at-a-time. The beginning is exciting and promising, but much more effort will be needed to take advantage of the huge potential that the new physics-based techniques offer.

Chromatin↗
PubMed2003

Assembly of single chromatin fibers depends on the tension in the DNA molecule: magnetic tweezers study.

We have used magnetic tweezers to study in real time chaperone-mediated chromatin assembly/disassembly at the level of single chromatin fibers. We find a strong dependence of the rate of assembly on the exerted force, with strong inhibition of assembly at forces exceeding 10 pN. During assembly, and especially at higher forces, occasional abrupt increases in the length of the fiber were observed, giving a clear indication of reversibility of the assembly process. This result is a clear demonstration of the dynamic equilibrium between nucleosome assembly and disassembly at the single chromatin fiber level.

Chromatin↗
PubMed2003

Magnetic tweezers: a sensitive tool to study DNA and chromatin at the single-molecule level.

The advent of single-molecule biology has allowed unprecedented insight into the dynamic behavior of biological macromolecules and their complexes. Unexpected properties, masked by the asynchronous behavior of myriads of molecules in bulk experiments, can be revealed; equally importantly, individual members of a molecular population often exhibit distinct features in their properties. Finally, the single-molecule approaches allow us to study the behavior of biological macromolecules under applied tension or torsion; understanding the mechanical properties of these molecules helps us understand how they function in the cell. In this review, we summarize the application of magnetic tweezers (MT) to the study of DNA behavior at the single-molecule level. MT can be conveniently used to stretch DNA and introduce controlled levels of superhelicity into the molecule and to follow to a high definition the action of different types of topoisomerases. Its potential for chromatin studies is also enormous, and we will briefly present our first chromatin results.

Biophysical Phenomena↗
PubMed2002

Stretching and imaging single DNA molecules and chromatin.

The advent of single-molecule biology has allowed unprecedented insight into the dynamic behavior of biological macromolecules and their complexes. Unexpected properties, masked by the asynchronous behavior of myriads of molecules in bulk experiments, can be revealed; equally importantly, individual members of a molecular population often exhibit distinct features in their properties. Finally, the single-molecule approaches allow us to study the behavior of biological macromolecules under applied tension or torsion: understanding the mechanical properties of these molecules helps us understand how they function in the cell. The aim of this chapter is to summarize and critically evaluate the properties of single DNA molecules and of single chromatin fibers. The use of the high-resolution imaging capabilities of the atomic force microscopy has been covered, together with manipulating techniques such as optical fibers, optical and magnetic tweezers, and flow fields. We have learned a lot about DNA and how it responds to applied forces. It is also clear that even though the study of the properties of individual chromatin fibers has just begun, the single-molecule approaches are expected to provide a wealth of information concerning the mechanical properties of chromatin and the way its structure changes during processes like transcription and replication.

Animals↗
PubMed2002

Single-molecule studies of chromatin fibers: a personal report.

With the advent of single-molecule techniques, macromolecular science has reached new horizons. Nowadays, we can observe, touch, stretch and twist biological macromolecules or their complexes, one-at-a time, in attempts to better understand their mechanical properties and to gain insights into their behavior in the living cell. Chromatin structure and function has been the focus of our research interests for many years. In the past decade, we have added some of the newly emerged single-molecule approaches to the more traditional biochemical and biophysical methods that we have been using throughout the years. This paper briefly summaries our studies on individual chromatin fibers using the atomic force microscope (AFM), optical tweezers, and magnetic tweezers. We believe that our results so far have contributed significantly to our understanding of chromatin, but we also hope that this is only the beginning, and that more exciting times lie ahead.

Biophysical Phenomena↗