Using computer simulations to model bacterial microcompartment assembly

Bacterial microcompartments are protein shells found in bacteria that surround enzymes and other chemicals required for certain biological reactions.  For example, the carboxysome is a type of microcompartment that enables bacteria to convert the products of photosynthesis into sugars (thus taking carbon out of the atmosphere).  During the formation of a microcompartment, the outer protein shell assembles around hundreds of enzymes and chemicals required for the reaction.  Because the intermediates in this assembly process are small and short-lived, it is hard to study in detail using experiments. It is therefore useful to develop computational models that can help explain how proteins collect the necessary cargo, and then assemble into an ordered shell with the cargo on the inside.  The videos in this post show some examples of computer simulations of a model for bacterial microcompartment assembly, with each video corresponding to a different set of parameters that control the strengths of interactions among the proteins and cargo.

The study is described in the research article “Many-molecule encapsulation by an icosahedral shell” by Jason Perlmutter, Farzaneh Mohajerani, and Michael Hagan in eLife (eLife 2016;10.7554/eLife.14078).

Video 1: Multistep assembly of a microcompartment encapsulating hundreds of molecules (I) video1
Video 2: Multistep assembly of a microcompartment encapsulating hundreds of molecules (II)  video2
Video 3: Assembly of a microcompartment and encapsulation of hundreds of diffuse cargo molecules  video3

Lipids hit a “sweet spot” to direct cellular membrane remodeling.

Lipid membrane reshaping is critical to many common cellular processes, including cargo trafficking, cell motility, and organelle biogenesis. The Rodal lab studies how dynamic membrane remodeling is achieved by the active interplay between lipids and proteins. Recent results, published in Cell Reports, demonstrate that for the membrane remodeling protein Nervous Wreck (Nwk), intramolecular autoregulation and membrane charge work together in surprising ways to restrict remodeling to a limited range of lipid compositions.

F-BAR (Fes/Cip4 homology Bin/Amphiphysin/Rvs) domain family proteins are important mediators of membrane remodeling events. The F-BAR domain forms a crescent-shaped α-helical dimer that interacts with and deforms negatively charged membrane phospholipids by assembling into higher-order scaffolds. In this paper, Kelley et al. have shown that the neuronal F-BAR protein Nwk is autoregulated by its C-terminal SH3 domains, which interact directly with the F-BAR domain to inhibit membrane binding. Until now, the dogma in the field has been that increasing concentrations of negatively charged lipids would increase Nwk membrane binding, and thus would induce membrane deformation.

Surprisingly, Kelley et al. found that autoregulation does not mediate this kind of simple “on-off” switch for membrane remodeling. Instead, increasing the concentration of negatively charged lipids increases membrane binding, but inhibits F-BAR membrane deforming activities (see below). Using a combination of in vitro assays and single particle electron microscopy, they found that the Nwk F-BAR domain efficiently assembles into higher-order structures and deforms membranes only within “sweet spot” of negative membrane charge, and that autoregulation elevates this range. The implication of this work is that autoregulation could either reduce membrane binding or promote higher-order assembly, depending on local cellular membrane composition. This study suggests a significant role for the regulation of membrane composition in remodeling.

Brandeis authors on the study included Molecular and Cell Biology graduate students Charlotte Kelley and Shiyu Wang, staff member Tania Eskin, and undergraduate Emily Messelaar ’13 from the Rodal lab; postdoctoral fellow Kangkang Song, Associate Professor of Biology Daniela Nicastro (currently at UT Southwestern), and Associate Professor of Physics Michael Hagan.

Kelley CF, Messelaar EM, Eskin TL, Wang S, Song K, Vishnia K, Becalska AN, Shupliakov O, Hagan MF, Danino D, Sokolova OS, Nicastro D, Rodal AA. Membrane Charge Directs the Outcome of F-BAR Domain Lipid Binding and Autoregulation. Cell reports. 2015;13(11):2597-609.

Simulations Say Viral Genome Lengths are Optimal for Capsid Assembly

Viruses are infectious agents made up of proteins and a genome made of DNA or RNA. Upon infecting a host cell, viruses hijack the cell’s gene expression machinery and force it to produce copies of the viral genome and proteins, which then assemble into new viruses that can eventually infect other host cells. Because assembly is an essential step in the viral life cycle, understanding how this process occurs could significantly advance the fight against viral diseases.

In many viral families, a protein shell called a capsid forms around the viral genome during the assembly process. Capsids can also assemble around nucleic acids in solution, indicating that a host cell is not required for their formation. Since capsid proteins are positively charged, and nucleic acids are negatively charged, electrostatic interactions between the two are thought to be important in capsid assembly. Current questions of interest are how structural features of the viral genome affect assembly, and why the negative charge on viral genomes is actually far greater than the positive charge on capsids. These questions are difficult to address experimentally because most of the intermediates that form during virus assembly are too short-lived to be imaged.

hagan-capsid-sim

Snapshots from a computer simulation in which model capsid subunits (blue) assemble around a linear, negatively charged polymer (red). Positive charges on the capsid proteins are shown in yellow.

In a new paper in eLife, Brandeis postdoc Jason Perlmutter, Physics grad student Cong Qiao, and Associate Professor Michael Hagan have used state of the art computational methods and advances in graphical processing units (on our High Performance Computing cluster) to produce the most realistic model of capsid assembly to date. They showed that the stability of the complex formed between the nucleic acid and the capsid depends on the length of the viral genome. Yield was highest for genomes within a certain range of lengths, and capsids that assembled around longer or shorter genomes tended to be malformed.

Perlmutter et al. also explored how structural features of the virus — including base-pairing between viral nucleic acids, and the size and charge of the capsid — determine the optimal length of the viral genome. When they included structural data from real viruses in their simulations and predicted the optimal lengths for the viral genome, the results were very similar to those seen in existing viruses. This indicates that the structure of the viral genome has been optimized to promote packaging into capsids. Understanding this relationship between structure and packaging will make it easier to develop antiviral agents that thwart or misdirect virus assembly, and could aid the redesign of viruses for use in gene therapy and drug delivery.

Perlmutter JD, Qiao C, Hagan MF. Viral genome structures are optimal for capsid assembly. eLife 2013;2:e00632

More science

We’ve all been busy this spring writing grants and teaching courses and doing research and graduating(!), so lots of publications snuck by that we didn’t comment on. Here’s a few I think that might be interesting to our readers.

  • From Chris Miller‘s lab, bacterial antiporters do act as “virtual proton efflux pumps”:
  • nsrv2Are ninja stars responsible for controlling actin disassembly? Seems like the Goode lab might think so.
    • Chaudhry F, Breitsprecher D, Little K, Sharov G, Sokolova O, Goode BL. Srv2/cyclase-associated protein forms hexameric shurikens that directly catalyze actin filament severing by cofilin. Mol Biol Cell. 2013;24(1):31-41.
  • What do you get from statistical mechanics of self-propelled particles? The Hagan and Baskaran groups team up to find out.
  • From John Lisman and Ole Jensen (PhD ’98), thoughts about what the theta and gamma rhythms in the brain encode
  • From Mike Marr‘s lab, studeies using genome-wide nascent sequencing to understand how transcrption bursting is controlled in eukaryotic cells
  • From the Lau and Sengupta labs, RNAi pathways contribute to long term plasticity in worms that have gone through the Dauer stage
    • Hall SE, Chirn GW, Lau NC, Sengupta P. RNAi pathways contribute to developmental history-dependent phenotypic plasticity in C. elegans. RNA. 2013;19(3):306-19.
  • Can nanofibers selectively disrupt cancer cell types? Early results from Bing Xu‘s group.
    • Kuang Y, Xu B. Disruption of the Dynamics of Microtubules and Selective Inhibition of Glioblastoma Cells by Nanofibers of Small Hydrophobic Molecules. Angew Chem Int Ed Engl. 2013.

Baskaran Wins NSF-CAREER award to pursue research on active fluids

Dr. Aparna Baskaran of the Physics Department has been awarded the prestigious CAREER grant from the National Science Foundation that is a highly competitive development grant for early career tenure track faculty members. This grant will fund the research ongoing in Dr. Baskaran’s group on dynamics in active materials. Active materials are a novel class of complex fluids that are driven out of equilibrium at the level of individual entities. Examples of such systems include bacterial suspensions, cytoskeletal filaments interacting with motor proteins and inanimate systems such as self-propelled phoretic colloidal particles. The theoretical challenge in understanding these systems lies in the fact that, unlike traditional materials, we no longer have the scaffold of equilibrium on which to base the theoretical framework.  At the practical front, these materials exhibit novel properties not seen in regular materials.  Further, they form the physical framework of biological systems  in that regulatory mechanisms modulate the mechanical properties of this material in response to environmental stimuli.  Dr. Baskaran’s research in this field will be done in collaboration with the groups of Dr. Michael Hagan, Dr. Zvonimir Dogic and Dr. Bulbul Chakraborty. It will enhance and complement the MRSEC research activities in the active materials thrust.

Figure Caption : Videos of example systems for active materials. A) A fish school exhibiting complex collective swimming. B) Swarming at the edge of an E. Coli Bacterial Colony. C) Cytoplasmic streaming inside the yolk of a fertilized cell.

Six scientists secure fellowships

One current undergraduate, and five alumni, from the Brandeis Sciences were honored with offers of National Science Foundation Graduate Research Fellowships in 2012. The fellowships, which are awarded based on a national competition, provide three full years of support for Ph.D. research and are highly valued by students and institutions. These students are:

  • Samuel McCandlish ’12 (Physics) , a current student who did research with Michael Hagan and Aparna Baskaran, resulting in a paper “Spontaneous segregation of self-propelled particles with different motilities” in Soft Matter (as a junior). He then switched to work with Albion Lawrence for his senior thesis research. Sam will speak about “Bending and Breaking Time Contours: a World Line Approach to Quantum Field Theory” at the Berko Symposium on May 14.  Sam has been offered a couple of other fellowships as well, so he’ll have a nice choice to make. Sam will be heading to Stanford in the fall to continue his studies in theoretical physics.
  • Briana Abrahms ’08 (Physics). After graduating from Brandeis, Briana followed her interests in ecological and conversation issues, and  in Africa as a research assistant with the Botswana Predator Conservation Trust, Briana previously described some of her experiences here in “Three Leopards and a Shower“. Briana plans to pursue as Ph.D. in Ecology at UC Davis.
  • Sarah Robinson ’07 (Chemistry). Sarah did undergraduate research with Irving Epstein on “Pattern formation in a coupled layer reaction-diffusion system”. After graduating, Sarah spent time with the Peace Corps in Tanzania, returning to study Neurosciene at UCSF.
  • Si Hui Pan ’10 (Physics) participated in a summer REU program at Harvard, and continued doing her honors thesis in collaboration with the labs at Harvard. Her award is to study condensed matter physics at MIT.
  • Elizabeth Setren ’10 was a Mathematics and Economics double major who worked together with Donald Shepard (Heller School) on the cost of hunger in the US. She has worked as an Assistant Economist at the Federal Reserve Bank of New York and her award is to study Economics at Harvard.
  • Michael Ari Cohen ’01 (Psychology) worked as a technology specialist for several years before returning to academia as  PhD student in the Energy and Resources Group at UC Berkeley.

Congratulations to all the winners!

Why nanorods assemble

In a recent paper in Phys. Rev. E, Brandeis postdoc Yasheng Yang and Assistant Professor of Physics Michael Hagan developed a theory to describe the assembly behavior of colloidal rods (i.e. nanorods) in the presence of inert polymer (which induces attractions between the nanorods). The nanorods assemble into several kinds of structures, including colloidal membranes, which  are two dimensional membrane-like structures composed of a one rod-length thick monolayer of aligned rods.  The theory shows that colloidal membranes are stabilized against stacking on top of each other by an entropic force arising from protrusions of rods from the membranes and that there is a critical aspect ratio (rod length/rod diameter) below which membranes are never stable. Understanding the forces that stabilize colloidal membranes is of practical importance since these structures could enable the manufacture of inexpensive and easily scalable optoelectronic devices. This work was part of a collaboration with the experimental lab of Zvonimir Dogic at Brandeis, where colloidal membranes are developed and studied.

Yang YS, Hagan MF. Theoretical calculation of the phase behavior of colloidal membranes. Phys Rev E. 2011;84(5).

Collective behaviors in active matter

Active matter is describes systems whose constituent elements consume energy and are thus out-of-equilibrium. Examples include flocks or herds of animals, collections of cells, and components of the cellular cytoskeleton. When these objects interact with each other, collective behavior can emerge that is unlike anything possible with an equilibrium system. The types of behaviors and the factors that control them however, remain incompletely understood. In a recent paper in Physical Review Letters, “Excitable patterns in active nematics“, Giomi and coworkers develop a continuum theoretical description motivated by recent experiments from the Dogic group at Brandeis in which microtubules (filamentous cytoskeletal molecules) and clusters of kinesin (a molecular motor) exhibit dramatic spatiotemporal fluctuations in density and alignment. Specifically, they consider a hydrodynamic description for density, flow, and nematic alignment. In contrast to previous theories of this type, the degree of nematic alignment is allowed to vary in space and time.  Remarkably, the theory predicts that the interplay between non-uniform nematic order, activity and flow results in spatially modulated relaxation oscillations, similar to those seen in excitable media and biological examples such as the cardiac cycle. At even higher activity the dynamics is chaotic and leads to large-scale swirling patterns which resemble those seen in recent experiments. An example of the flow pattern is shown below left, and the nematic order parameter, which describes the degree of alignment of the filaments, as shown for the same configuration below right. These predictions can be tested in future experiments on systems of microtubules and motor proteins.

The system behavior for an active nematic at high activity. (left) The velocity field (arrows) is superimposed on a plot of the concentration of active nematogens (green=large concentration, red=small concentration). (right) A plot of the nematic order parameter, S,  (blue=large S, brown=small S) is superimposed on a plot of the nematic director (arrows). The flow under high activity is characterized by large vortices that span lengths of the order of the system size and the director field is organized in grains.

 

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