Esther Bullitt, Ph.D.
A.B. Grinnell College
|Phone: (617) 638-5037
Fax: (617) 638-4041
Address: see below
Link to BU Faculty Profile
Link to ORCID
Protein Structure Facilitates Function: Using Electron Microscopy and Quantitative Image Analysis to see how Macromolecular Assemblies Work
Structural studies of biological macromolecular assemblies are providing an understanding of cellular function. In our laboratory, we utilize electron microscopy and image reconstruction to investigate questions about how adhesion pili aid pathogenic bacterial survival under harsh physiological conditions.
TYPE III SECRETION
Only 10 bacteria (or fewer!) are needed for Shigella flexneri to cause dysentery (bloody diarrhea). This disease is initiated via the type III secretion system, which is used to secrete both toxins and bacterial effectors that alter normal host cell functions to promote bacterial growth and spread.
To understand (and disrupt) the process of infection, we are examining assembly intermediates of the T3SS syringe-like needle and its tip structure. Through our collaborations with the Picking lab at Oklahoma State University Stillwater and the Geisbrecht lab at the University of Missouri Kansas City we have shown that IpaD, the first protein that localizes to the needle tip, is present as a pentamer, and undergoes a dramatic conformational change as compared to the structure that has been solved by x-ray crystallography.
The structures of three-dimensional reconstructions of immature and nascent needle tips show clearly IpaD as an elongated pentameric structure.
This new result demonstrates that the distal domain of IpaD flips up, as compared to the structure solved by X-ray crystallography (pdb 2j0o).
Diagram to the right shows the conformational change of the distal domain that is needed to fit the crystal structure of IpaD into our density map of 3-dimensional reconstructions from electron microscopy data.
Epler, C.R., N.E. Dickenson, E. Bullitt*, W.L. Picking* (2012). Ultrastructural analysis of IpaD at the tip of the nascent MxiH type III secretion apparatus of Shigella flexneri. J. Mol. Biol. 420:29-39. PMID: 22480614 *corresponding authors
The poliovirus polymerase, 3Dpol, is an RNA-dependent RNA polymerase. Its structure is similar to other polymerases, and can be described as a right hand. We are investigating the role of two-dimensional 3Dpol lattices in replication, using electron microscopy and image processing of tubes and sheets formed in vitro. The crystal structure shown was solved in Steve Schultz’s lab, pdb 1rdr.
In collaboration with Dr. Karla Kirkegaard’s lab at Stanford University we have shown that
1) 3Dpol forms two-dimensional lattices that we expect facilitate replication by concentrating substrate and enzyme onto a two-dimensional surface, the vesicular membrane,
2) “Zombie” 3Dpol protein (dead active site) can support assembly of oligomers and restore replication when the wild type protein concentration is too low to do so.
Lyle, J.M.*, E. Bullitt*, K. Bienz, K. Kirkegaard (2002). Visualization and functional analysis of RNA-dependent RNA polymerase lattices. Science 296:2218-2222. *these authors contributed equally
Spagnolo, J.F., E. Rossignol, E. Bullitt, K. Kirkegaard (2010). Enzymatic and non-enzymatic functions of viral RNA-dependent RNA polymerases within oligomeric arrays. RNA 16:382-393. PMCID: PMC2811667
Tellez, A.B*., J. Wang*, E.J. Tanner, J.F. Spagnolo, K. Kirkegaard+ and E. Bullitt+ (2011). “Interstitial Contacts in an RNA-Dependent RNA Polymerase Lattice.” J Mol Biol 412(4): 737-50. PMID: 21839092. http://www.ncbi.nlm.nih.gov/pubmed/21839092.
*these authors contributed equally, +corresponding authors
BACTERIAL ADHESION PILI
Fibers on the surfaces of many pathogenic bacteria can overextend like a toy Slinky, whereas homologous proteins on bacteria that cause pneumonia, ear infections, and meningitis assemble into a 3-stranded rope-like fibers (blue). In collaboration with Dr. Magnus Andersson’s lab at Umeå University in Umeå Sweden we have shown that the forces required for unwinding pili (also called ‘fimbriae’) are specific for the pilus-type, and thus vary with the microenvironment expected to be encountered. For example, CFA/I pili on diarrhea-causing intestinal bacteria unwind at less than one-third the force required to unwind P-pili expressed on urinary tract infection bacteria.
Our structural studies have shown that the mechanisms by which pili assemble on the bacterial outer membane include a loop region that allows rotation into a helical filament (P-pili) and, in collaboration with Capt. Stephen Savarino’s lab at the Naval Medical Reseach Center in Silver Spring, a proline trans to cis isomeration (CFA/I pili).
As more bacteria become resistant to antibiotics, it is essential to develop novel approaches for prevention and treatment of infection. Structural studies are vital for discovering clues to how bacteria bind, and how they remain attached while the host is trying to remove them. We are examining the architecture of bacterial adhesion pili and investigating small molecules that disrupt their assembly and/or function.
Bullitt, E., L. Makowski (1995). Structural polymorphism of bacterial adhesion pili. Nature 373:164-167. PMID: 7816100
Mu, X.Q. and E. Bullitt (2006). Structure and assembly of P-pili: a protruding hinge region used for assembly of a bacterial adhesion filament. Proc. Natl. Acad. Sci. USA 103:9861-9866. PMCID: PMC1502544
Mu, X.Q., S.J. Savarino, E. Bullitt (2008). The three-dimensional structure of CFA/I adhesion pili: Traveler’s Diarrhea Bacteria Hang on by a Spring. J. Mol. Biol. 376:614-620. PMCID: PMC2265596
Li, Y-F, S. Poole, K. Nishio, K. Jang, F. Rasulova, A. McVeigh, S.J. Savarino, D. Xia, E. Bullitt (2009) Structure of CFA/I fimbriae from enterotoxigenic Escherichia coli. Proc. Natl. Acad. Sci. USA 106: 10793-10798. PMCID: PMC2705562
Andersson M, O. Björnham, M. Svantesson, A. Badahdah, B.E. Uhlin, E. Bullitt (2012). A Structural Basis for Sustained Bacterial Adhesion: Biomechanical Properties of CFA/I Pili. J. Mo.l Biol. 415: 918-928. PMCID: PMC3267891
Andersson M, Björnham O, Svantesson M, Badahdah A, Uhlin BE, Bullitt E. (2012). A Structural Basis for Sustained Bacterial Adhesion: Biomechanical Properties of CFA/I Pili., J Mol Biol. 2012 Feb 3;415(5):918-28.
Tellez AB, Wang J, Tanner EJ, Spagnolo JF, Kirkegaard K, Bullitt E. (2011). Interstitial contacts in an RNA-dependent RNA polymerase lattice. J Mol Biol. 2011 Sep 30;412(4):737-50.
Li, Y-F, S. Poole, K. Nishio, K. Jang, F. Rasulova, A. McVeigh, S.J. Savarino, D. Xia, E. Bullitt (2009). Structure of CFA/I fimbriae from enterotoxigenic Escherichia coli. Proc. Natl. Acad. Sci. 106:10793-10798.
Mu X.Q., S.J. Savarino, E. Bullitt (2008). The three-dimensional structurel of CFA/I adhesion pili: Traveler’s diarrhea bacteria hang on by a spring. J. Mol. Biol. 376:614-620.
Verger, D., E. Bullitt, S.J. Hultgren, G. Waksman (2007). Crystal structure of the P pilus rod subunit PapA. PLoS Pathogens 3:e73(674-682).
Mu XQ and E. Bullitt (2006). Structure and assembly of P-pili: a protruding hinge region used for assembly of a bacterial adhesion filament. Proc. Natl. Acad. Sci.103:9861-9866.
Mu X.Q., E.H. Egelman, E. Bullitt (2002). Structure and Function of Hib Pili from Haemophilus influenzae Type b. J Bacteriol. 184:4868-74.
Lyle J.M., E. Bullitt, K. Bienz, K. Kirkegaard (2002). Visualization and functional analysis of RNA-dependent RNA polymerase lattices. Science. Jun 21;296(5576):2218-22.
Bullitt E., M.P. Rout, J.V. Kilmartin, C.W. Akey(1997). The yeast spindle pole body is assembled around a central crystal of Spc42p. Cell Jun 27;89(7):1077-86.
Bullitt E, L. Makowski (1995). Structural polymorphism of bacterial adhesion pili. Nature Jan 12;373(6510):164-7.
Department of Physiology and Biophysics
Boston University School of Medicine
700 Albany Street
Boston MA 02118-2526
Phone: (617) 638-5037
Fax: (617) 638-4041