5,812 research outputs found
First-principles calculation of DNA looping in tethered particle experiments
We calculate the probability of DNA loop formation mediated by regulatory
proteins such as Lac repressor (LacI), using a mathematical model of DNA
elasticity. Our model is adapted to calculating quantities directly observable
in Tethered Particle Motion (TPM) experiments, and it accounts for all the
entropic forces present in such experiments. Our model has no free parameters;
it characterizes DNA elasticity using information obtained in other kinds of
experiments. [...] We show how to compute both the "looping J factor" (or
equivalently, the looping free energy) for various DNA construct geometries and
LacI concentrations, as well as the detailed probability density function of
bead excursions. We also show how to extract the same quantities from recent
experimental data on tethered particle motion, and then compare to our model's
predictions. [...] Our model successfully reproduces the detailed distributions
of bead excursion, including their surprising three-peak structure, without any
fit parameters and without invoking any alternative conformation of the LacI
tetramer. Indeed, the model qualitatively reproduces the observed dependence of
these distributions on tether length (e.g., phasing) and on LacI concentration
(titration). However, for short DNA loops (around 95 basepairs) the experiments
show more looping than is predicted by the harmonic-elasticity model, echoing
other recent experimental results. Because the experiments we study are done in
vitro, this anomalously high looping cannot be rationalized as resulting from
the presence of DNA-bending proteins or other cellular machinery. We also show
that it is unlikely to be the result of a hypothetical "open" conformation of
the LacI tetramer.Comment: See the supplement at
http://www.physics.upenn.edu/~pcn/Ms/TowlesEtalSuppl.pdf . This revised
version accepted for publication at Physical Biolog
Concentration and Length Dependence of DNA Looping in Transcriptional Regulation
In many cases, transcriptional regulation involves the binding of transcription factors at sites on the DNA that are not immediately adjacent to the promoter of interest. This action at a distance is often mediated by the formation of DNA loops: Binding at two or more sites on the DNA results in the formation of a loop, which can bring the transcription factor into the immediate neighborhood of the relevant promoter. These processes are important in settings ranging from the historic bacterial examples (bacterial metabolism and the lytic-lysogeny decision in bacteriophage), to the modern concept of gene regulation to regulatory processes central to pattern formation during development of multicellular organisms. Though there have been a variety of insights into the combinatorial aspects of transcriptional control, the mechanism of DNA looping as an agent of combinatorial control in both prokaryotes and eukaryotes remains unclear. We use single-molecule techniques to dissect DNA looping in the lac operon. In particular, we measure the propensity for DNA looping by the Lac repressor as a function of the concentration of repressor protein and as a function of the distance between repressor binding sites. As with earlier single-molecule studies, we find (at least) two distinct looped states and demonstrate that the presence of these two states depends both upon the concentration of repressor protein and the distance between the two repressor binding sites. We find that loops form even at interoperator spacings considerably shorter than the DNA persistence length, without the intervention of any other proteins to prebend the DNA. The concentration measurements also permit us to use a simple statistical mechanical model of DNA loop formation to determine the free energy of DNA looping, or equivalently, the J-factor for looping
Structure-based stabilization of insulin as a therapeutic protein assembly via enhanced aromatic-aromatic interactions
Key contributions to protein structure and stability are provided by weakly polar interactions, which arise from asymmetric electronic distributions within amino acids and peptide bonds. Of particular interest are aromatic side chains whose directional π-systems commonly stabilize protein interiors and interfaces. Here, we consider aromatic-aromatic interactions within a model protein assembly: the dimer interface of insulin. Semi-classical simulations of aromatic-aromatic interactions at this interface suggested that substitution of residue TyrB26 by Trp would preserve native structure while enhancing dimerization (and hence hexamer stability). The crystal structure of a [TrpB26]insulin analog (determined as a T3Rf3 zinc hexamer at a resolution of 2.25 Å) was observed to be essentially identical to that of WT insulin. Remarkably and yet in general accordance with theoretical expectations, spectroscopic studies demonstrated a 150-fold increase in the in vitro lifetime of the variant hexamer, a critical pharmacokinetic parameter influencing design of long-acting formulations. Functional studies in diabetic rats indeed revealed prolonged action following subcutaneous injection. The potency of the TrpB26-modified analog was equal to or greater than an unmodified control. Thus, exploiting a general quantum-chemical feature of protein structure and stability, our results exemplify a mechanism-based approach to the optimization of a therapeutic protein assembly
The Mechanics and Statistics of Active Matter
Active particles contain internal degrees of freedom with the ability to take
in and dissipate energy and, in the process, execute systematic movement.
Examples include all living organisms and their motile constituents such as
molecular motors. This article reviews recent progress in applying the
principles of nonequilibrium statistical mechanics and hydrodynamics to form a
systematic theory of the behaviour of collections of active particles -- active
matter -- with only minimal regard to microscopic details. A unified view of
the many kinds of active matter is presented, encompassing not only living
systems but inanimate analogues. Theory and experiment are discussed side by
side.Comment: This review is to appear in volume 1 of the Annual Review of
Condensed Matter Physics in July 2010 and is posted here with permission from
that journa
An Assessment of a 15 vs. 30 Second Recovery Period on Vertical Jump Performance
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Assessing the Impact of a Governed Focal Point on Broad Jump Performance in Collegiate Females
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Exact theory of kinkable elastic polymers
The importance of nonlinearities in material constitutive relations has long
been appreciated in the continuum mechanics of macroscopic rods. Although the
moment (torque) response to bending is almost universally linear for small
deflection angles, many rod systems exhibit a high-curvature softening. The
signature behavior of these rod systems is a kinking transition in which the
bending is localized. Recent DNA cyclization experiments by Cloutier and Widom
have offered evidence that the linear-elastic bending theory fails to describe
the high-curvature mechanics of DNA. Motivated by this recent experimental
work, we develop a simple and exact theory of the statistical mechanics of
linear-elastic polymer chains that can undergo a kinking transition. We
characterize the kinking behavior with a single parameter and show that the
resulting theory reproduces both the low-curvature linear-elastic behavior
which is already well described by the Wormlike Chain model, as well as the
high-curvature softening observed in recent cyclization experiments.Comment: Revised for PRE. 40 pages, 12 figure
Covariation between oxygen and hydrogen stable isotopes declines along the path from xylem water to wood cellulose across an aridity gradient
Oxygen and hydrogen isotopes of cellulose in plant biology are commonly used to infer environmental conditions, often from time series measurements of tree rings. However, the covariation (or the lack thereof) between δ18O and δ2H in plant cellulose is still poorly understood. We compared plant water, and leaf and branch cellulose from dominant tree species across an aridity gradient in Northern Australia, to examine how δ18O and δ2H relate to each other and to mean annual precipitation (MAP). We identified a decline in covariation from xylem to leaf water, and onwards from leaf to branch wood cellulose. Covariation in leaf water isotopic enrichment (Δ) was partially preserved in leaf cellulose but not branch wood cellulose. Furthermore, whilst δ2H was well-correlated between leaf and branch, there was an offset in δ18O between organs that increased with decreasing MAP. Our findings strongly suggest that postphotosynthetic isotope exchange with water is more apparent for oxygen isotopes, whereas variable kinetic and nonequilibrium isotope effects add complexity to interpreting metabolic-induced δ2H patterns. Varying oxygen isotope exchange in wood and leaf cellulose must be accounted for when δ18O is used to reconstruct climatic scenarios. Conversely, comparing δ2H and δ18O patterns may reveal environmentally induced shifts in metabolism
NEXUS/Physics: An interdisciplinary repurposing of physics for biologists
In response to increasing calls for the reform of the undergraduate science
curriculum for life science majors and pre-medical students (Bio2010,
Scientific Foundations for Future Physicians, Vision & Change), an
interdisciplinary team has created NEXUS/Physics: a repurposing of an
introductory physics curriculum for the life sciences. The curriculum interacts
strongly and supportively with introductory biology and chemistry courses taken
by life sciences students, with the goal of helping students build general,
multi-discipline scientific competencies. In order to do this, our two-semester
NEXUS/Physics course sequence is positioned as a second year course so students
will have had some exposure to basic concepts in biology and chemistry.
NEXUS/Physics stresses interdisciplinary examples and the content differs
markedly from traditional introductory physics to facilitate this. It extends
the discussion of energy to include interatomic potentials and chemical
reactions, the discussion of thermodynamics to include enthalpy and Gibbs free
energy, and includes a serious discussion of random vs. coherent motion
including diffusion. The development of instructional materials is coordinated
with careful education research. Both the new content and the results of the
research are described in a series of papers for which this paper serves as an
overview and context.Comment: 12 page
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