Research Focus
Honors
Selected Publications
Contact
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Hughson?Group Webpage

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Faculty Assistant
Anna Schmedel
schedel@princeton.edu
Phone: 609-258-5028



Research Focus

Intracellular Trafficking

One major focus of the research in my group is to understand the protein machinery that generates the interior architecture of cells by guiding the movement and fusion of intracellular transport vesicles. All eukaryotic cells contain a profusion of membrane-bounded compartments. Traffic among these compartments is brisk. Cargo is transported in membrane vesicles that bud from one compartment, travel through the cell, and deliver their contents by fusing with another compartment. Underlying this intricate choreography is a set of proteins and protein complexes responsible for the creation, movement, docking, and fusion of vesicles. Our lab seeks to understand the design principles that endow these protein nanomachines with the ability to manipulate membrane vesicles, thereby powering a bustling intracellular transportation network.
Several of our current projects involve structural and mechanistic studies of large multi-subunit protein complexes that orchestrate the docking and fusion of transport vesicles. These complexes guide cargo-laden vesicles to their destinations and coordinate the activities of other components of the trafficking machinery, including the 'SNARE' proteins that catalyze membrane fusion itself. We are investigating the structure and function of these complexes using an array of technologies including x-ray crystallography, electron microscopy, site-directed mutagenesis, in vitro reconstitution, and a suite of spectroscopic techniques.

The initial recognition between a vesicle and its membrane target, on the other hand, is mediated by large protein complexes called tethering factors. Tethering factors act upstream of SNARE complex assembly and play key roles in determining the specificity of trafficking. Because little is known about the structures of tethering factors or the mechanism(s) by which they act, we have recently initiated biochemical and biophysical studies of these key protein complexes.

Quorum Sensing

A second major focus within our group is bacteria, and the remarkable finding that these single-celled organisms communicate with one another by emitting and receiving small-molecule signals. We are interested in cataloging the signal molecules and in understanding their biosynthesis and detection. Moreover, since bacteria often respond to these signals in undesirable ways – like forming antibiotic-resistant biofilms or mounting an attack on a human host – we are interested in discovering and characterizing molecules that interfere with bacterial communication. We are investigating these issues using a range of biochemical and biophysical approaches. For example, we are attempting to determine the crystal structures of the enzymes responsible for synthesizing the signal molecules, and of the receptors responsible for detecting them.

We have also begun to identify antagonists – molecules that inhibit signaling – and we are interested in using biochemical and structural methods to figure out how they work and how they might be improved by rational design. Finally, in collaboration with other Princeton labs in the molecular biology, chemistry, and physics departments, we are trying to understand how bacteria integrate the information they receive via multiple different signal molecules to craft an appropriate response.


Research Areas
Chemical Biology



Honors
Advisory Board,PLoSBiology (2012-present)
Clare Hall Life Member, Cambridge UK (2007-present)
Beckman Young Investigator (1995-1997)
SearleScholar (1995-1998)


Selected Recent Publications
Baker, R. W.; Jeffrey, P. D.; Hughson, F. M., "Crystal Structures of the Sec1/Munc18 (SM) Protein Vps33, Alone and Bound to the Homotypic Fusion and Vacuolar Protein Sorting (HOPS) Subunit Vps16."Plos One 2013, 8 (6).
Bharucha, N.; Liu, Y.; Papanikou, E.; McMahon, C.; Esaki, M.; Jeffrey, P. D.; Hughson, F. M.; Glick, B. S., "Sec16 influences transitional ER sites by regulating rather than organizing COPII."Molecular Biology of the Cell 2013, 24 (21), 3406-3419.
McMahon, C.;Studer, S. M.;Clendinen, C.;Dann, G. P.; Jeffrey, P. D.;Hughson, F. M., "The Structure ofSec12Implicates Potassium Ion Coordination inSar1Activation."Journal of Biological Chemistry2012,287(52), 43599-43606.
Cavanaugh, L. F.; Chen, X.; Richardson, B. C.; Ungar, D.; Pelczer, I.; Rizo, J.; Hughson, F. M., "Structural analysis of conserved oligomeric golgi complex subunit 2."Journal of Biological Chemistry 2007, 282 (32), 23418-23426.
Chen, G.; Swem, L. R.; Swem, D. L.; Stauff, D. L.; O'Loughlin, C. T.; Jeffrey, P. D.; Bassler, B. L.; Hughson, F. M., "A Strategy for Antagonizing Quorum Sensing."Molecular Cell 2011, 42 (2), 199-209.
Yen, W.-L.;Shintani, T.;Nair, U.; Cao, Y.; Richardson, B. C.; Li, Z.;Hughson, F. M.; Baba, M.;Klionsky, D. J., "The conservedoligomericGolgi complex is involved in double-membrane vesicleformationduring autophagy."Journal of Cell Biology2010,188(1), 101-114.
Hughson, F. M.; Reinisch, K. M., "Structure and mechanism in membrane trafficking."Current Opinion in Cell Biology 2010, 22 (4), 454-460.
Lees, J. A.; Yip, C. K.;Walz, T.;Hughson, F. M., "Molecular organization of the COG vesicle tethering complex."Nature Structural & Molecular Biology2010,17(11),1292-U139.
Ren, Q.;Wimmer, C.;Chicka, M. C.; Ye, S.; Ren, Y.;Hughson, F. M.;Whiteheart, S. W., "Munc13-4is a limiting factor in the pathway required for platelet granule release and hemostasis."Blood2010,116(6), 869-877.
Ren, Y.; Yip, C. K.;Tripathi, A.;Huie, D.; Jeffrey, P. D.;Walz, T.;Hughson, F. M., "A Structure-Based Mechanism for Vesicle Capture by theMultisubunitTethering Complex Dsl1."Cell2009,139(6), 1119-1129.
Richardson, B. C.; Smith, R. D.;Ungar, D.;Nakamura, A.; Jeffrey, P. D.;Lupashin, V. V.;Hughson, F. M., "Structural basis for a humanglycosylationdisorder caused by mutation of theCOG4gene."Proceedings of the National Academy of Sciences of the United States of America2009,106(32), 13329-13334.
Kelly, R. C.; Bolitho, M. E.; Higgins, D. A.; Lu, W.; Ng, W.-L.; Jeffrey, P. D.; Rabinowitz, J. D.; Semmelhack, M. F.; Hughson, F. M.; Bassler, B. L., "The Vibrio cholerae quorum-sensing autoinducer CAI-1: analysis of the biosynthetic enzyme CqsA."Nature Chemical Biology 2009, 5 (12), 891-895.
Tripathi, A.; Ren, Y.; Jeffrey, P. D.;Hughson, F. M., "Structural characterization ofTip20pandDsl1p, subunits of theDsl1pvesicle tethering complex."Nature Structural & Molecular Biology2009,16(2), 114-123.
Neiditch, M. B.;Federle, M. J.;Pompeani, A. J.; Kelly, R. C.;Swem, D. L.; Jeffrey, P. D.;Bassler, B. L.;Hughson, F. M., "Ligand-inducedasymmetry inhistidinesensorkinasecomplex regulates quorum sensing."Cell2006,126(6), 1095-1108.
Togneri, J.; Cheng, Y.-S.; Munson, M.;Hughson, F. M.; Carr, C. M., "Specific SNARE complex binding mode of theSec1/Munc-18protein, Sec1p."Proceedings of the National AcademyofSciences of the United States of America2006,103(47), 17730-17735.
Ungar, D.; Oka, T.;Vasile, E.;Krieger, M.;Hughson, F. M., "Subunit architecture of the conservedoligomericgolgicomplex."Journal of Biological Chemistry2005,280(38), 32729-32735.
Neiditch, M. B.;Federle, M. J.; Miller, S. T.;Bassler, B. L.;Hughson, F. M., "Regulation ofLuxPQreceptor activity by the quorum-sensing signal autoinducer-2."Molecular Cell2005,18(5), 507-518.
Oka, T.;Vasile, E.; Penman, M.;Novina, C. D.;Dykxhoorn, D. M.;Ungar, D.;Hughson, F. M.;Krieger, M., "Genetic analysis of the subunit organization and functionof the conservedoligomericgolgi(COG) complex - Studies of Cog5- andCog7-deficientmammalian cells."Journal of Biological Chemistry2005,280(38), 32736-32745.
Miller, S. T.; Xavier, K. B.; Campagna, S. R.; Taga, M. E.; Semmelhack, M. F.; Bassler, B. L.; "Hughson, F. M., Salmonella typhimurium recognizes a chemically distinct form of the bacterial quorum-sensing signal Al-2."Molecular Cell 2004, 15 (5), 677-687.
Zweifel, M. E.; Leahy, D. J.;Hughson, F. M.;Barrick, D., "Structure and stability of theankyrindomain of theDrosophilaNotch receptor."Protein Science2003,12(11), 2622-2632.
Munson, M.; Hughson, F. M., "Conformational regulation of SNARE assembly and disassembly in vivo."Journal of Biological Chemistry 2002, 277 (11), 9375-9381.
Chen, X.;Schauder, S.;Potier, N.; VanDorsselaer, A.;Pelczer, I.;Bassler, B. L.;Hughson, F. M., "Structural identification of a bacterial quorum-sensing signal containing boron."Nature2002,415(6871), 545-549.
Lerman, J. C.;Robblee, J.;Fairman, R.;Hughson, F. M., "Structural analysis of the neuronal SNARE protein syntaxin-1A."Biochemistry2000,39(29), 8470-8479.
Waters, M. G.;Hughson, F. M., "Membrane tethering and fusion in the secretory andendocyticpathways."Traffic2000,1(8), 588-597.
Munson, M.; Chen, X.;Cocina, A. E.; Schultz, S. M.;Hughson, F. M., "Interactions within the yeast t-SNARESso1pthat control SNARE complex assembly."Nature Structural Biology2000,7(10), 894-902.