SYNERGY OF DNA-BENDING NUCLEOID PROTEINS AND MACROMOLECULAR CROWDING IN CONDENSING DNA

2007 ◽  
Vol 02 (03n04) ◽  
pp. 259-265 ◽  
Author(s):  
ESIO BESSA RAMOS ◽  
KATHELIJNE WINTRAECKEN ◽  
ANS GEERLING ◽  
RENKO DE VRIES

Many prokaryotic nucleoid proteins bend DNA and form extended helical protein-DNA fibers rather than condensed structures. On the other hand, it is known that such proteins (such as bacterial HU) strongly promote DNA condensation by macromolecular crowding. Using theoretical arguments, we show that this synergy is a simple consequence of the larger diameter and lower net charge density of the protein-DNA filaments as compared to naked DNA, and hence, should be quite general. To illustrate this generality, we use light-scattering to show that the 7kDa basic archaeal nucleoid protein Sso7d from Sulfolobus solfataricus (known to sharply bend DNA) likewise does not significantly condense DNA by itself. However, the resulting protein-DNA fibers are again highly susceptible to crowding-induced condensation. Clearly, if DNA-bending nucleoid proteins fail to condense DNA in dilute solution, this does not mean that they do not contribute to DNA condensation in the context of the crowded living cell.

2005 ◽  
Vol 35 (3) ◽  
pp. 214-223 ◽  
Author(s):  
David Pastré ◽  
Olivier Piétrement ◽  
Fabrice Landousy ◽  
Loïc Hamon ◽  
Isabelle Sorel ◽  
...  

Author(s):  
William Krakow ◽  
Benjamin Siegel

Unwin has used a metallized non-conducting thread in the back focal plane of the objective lens that stops out a portion of the unscattered beam, takes on a localized positive charge and thus produces an additional phase shift to give a different transfer function of the lens. Under the particular conditions Unwin used, the phase contrast image was shifted to bright phase contrast for optimum focus.We have investigated the characteristics of this type of electrostatic phase plate, both analytically and experimentally, as functions of the magnitude of charge and defocus. Phase plates have been constructed by using Wollaston wire to mount 0.25μ diameter platinum wires across apertures ranging from 50 to 200μ diameter and vapor depositing SiO and gold on the mounted wires to give them the desired charging characteristics. The net charge was varied by adjusting only the bias on the Wehnelt shield of the gun, and hence the beam currents and effective size of the source.


Author(s):  
P. Pradère ◽  
J.F. Revol ◽  
R. St. John Manley

Although radiation damage is the limiting factor in HREM of polymers, new techniques based on low dose imaging at low magnification have permitted lattice images to be obtained from very radiation sensitive polymers such as polyethylene (PE). This paper describes the computer averaging of P4MP1 lattice images. P4MP1 is even more sensitive than PE (total end point dose of 27 C m-2 as compared to 100 C m-2 for PE at 120 kV). It does, however, have the advantage of forming flat crystals from dilute solution and no change in d-spacings is observed during irradiation.Crystals of P4MP1 were grown at 60°C in xylene (polymer concentration 0.05%). Electron microscopy was performed with a Philips EM 400 T microscope equipped with a Low Dose Unit and operated at 120 kV. Imaging conditions were the same as already described elsewhere. Enlarged micrographs were digitized and processed with the Spider image processing system.


Author(s):  
Yimei Zhu ◽  
J. Tafto

The electron holes confined to the CuO2-plane are the charge carriers in high-temperature superconductors, and thus, the distribution of charge plays a key role in determining their superconducting properties. While it has been known for a long time that in principle, electron diffraction at low angles is very sensitive to charge transfer, we, for the first time, show that under a proper TEM imaging condition, it is possible to directly image charge in crystals with a large unit cell. We apply this new way of studying charge distribution to the technologically important Bi2Sr2Ca1Cu2O8+δ superconductors.Charged particles interact with the electrostatic potential, and thus, for small scattering angles, the incident particle sees a nuclei that is screened by the electron cloud. Hence, the scattering amplitude mainly is determined by the net charge of the ion. Comparing with the high Z neutral Bi atom, we note that the scattering amplitude of the hole or an electron is larger at small scattering angles. This is in stark contrast to the displacements which contribute negligibly to the electron diffraction pattern at small angles because of the short g-vectors.


Author(s):  
Sengshiu Chung ◽  
Peggy Cebe

We are studying the crystallization and annealing behavior of high performance polymers, like poly(p-pheny1ene sulfide) PPS, and poly-(etheretherketone), PEEK. Our purpose is to determine whether PPS, which is similar in many ways to PEEK, undergoes reorganization during annealing. In an effort to address the issue of reorganization, we are studying solution grown single crystals of PPS as model materials.Observation of solution grown PPS crystals has been reported. Even from dilute solution, embrionic spherulites and aggregates were formed. We observe that these morphologies result when solutions containing uncrystallized polymer are cooled. To obtain samples of uniform single crystals, we have used two-stage self seeding and solution replacement techniques.


1979 ◽  
Vol 42 (05) ◽  
pp. 1452-1459 ◽  
Author(s):  
Robert H Yue ◽  
Toby Starr ◽  
Menard M Gertler

SummaryCommercial porcine heparin can be separated into three distinct subtractions by using DEAE-cellulose chromatography and a stepped salt gradient. Gram quantities of heparin can be fractionated by this technique. All three heparin subtractions can accelerate the inhibition of thrombin by antithrombin III with different efficiency. The specific activities of the high activity heparin, intermediate activity heparin and low activity heparin are 228 units/mg, 142 units/mg and 95 units/mg, respectively. Both the uronic acid content and the quantity of N-SO4 for all three heparin subfractions have been evaluated. The high activity heparin has the lowest uronic acid and N-SO4 content. The successful separation of commercial heparin into three distinct subfractions by means of ion-exchange chromatography suggests that the net charge on these three heparin components will serve as a model system in the elucidation of the structure and activity relationship to the biological function of heparin.


2003 ◽  
Author(s):  
Charles Thomas Parker ◽  
Dorothea Taylor ◽  
George M Garrity

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