Sacha De Carlo, PhD

Assistant Professor Chemistry Dept.

B.Sc. (1998, Biology & 1999, Physics), University of Lausanne, Switzerland.

Ph.D. (2002, Biophysics), University of Lausanne, Switzerland (Mentor: J. Dubochet).

Post-doctoral training: University of California at Berkeley (Mentor: E. Nogales) and University of Colorado at Boulder (Mentor: A. Hoenger).

Marshak Science Building - Office 1335 ; Lab 1321 Phone: Office, (212) 650-6070; Lab (212) 650-6582 Fax: (212) 650-6107 E-mail: sdecarlo[at]ccny.cuny.edu Lab URL: De Carlo Lab - 3D electron microscopy of molecular machines Doctoral Faculty for the PhD program in Chemistry/Biology/Biochemistry.

CUNY Institute for Macro Molecular Assemblies - CUNY MMA

Supported in part by the Research Centers in Minority Institutions (NIH/NCRR/RCMI) CCNY/Grant G12-RR03060.

People in the group

Dr. Jorge Morales - Electron Microscopy Facility Manager - Phone: x8591

Ms. Reman Daglawi - Biochemistry M.S. student - Phone: x6582

Mr. Chris Reyes - Biochemistry M.S. student - Phone: x6582

Undergraduate students are encouraged to apply for a PhD, as two positions are available immediately. The research involves routine molecular biology techniques, biochemistry and structural biology. We prepare complexes of molecular assemblies (see Research - below), isolate and purify them by chormatographic methods, and study them using exciting techniques such as cryo-electron microscopy. From images collected in the electron microscope we extract structural data that we process with the computer. Computer-assisted image processing is used to build 3D models of our structures and help us interpret the data and understand molecular mechanisms. As you can read, there are many exciting things you will learn in the DeCarlo lab!

In addition to the projects described here below, Prof De Carlo also collaborates with other Faculty at CCNY. A project is ongoing to study the structure of Influenza viruses (A & B) and their surface receptors by cryo-electron tomography, this is in collaboration with Prof Paul Gottlieb (Sophie Davis Medical School).

Nanomaterials are also studied by cryo-electron microscopy in an ongoing collaboration with Prof George John in the Chemistry Department.

Biophysics; Structural Biology; Biochemistry; Cryo-Electron Microscopy; 3-D Image Reconstruction Techniques

Recent work has focused on elucidating transcription initiation/elongation and regulatory mechanisms in Archaea (with D. Taatjes, Univ. of Colorado). Our results directly map the position of the initiation ‘core’ complex (TBP/TFB transcription factors) onto the archaeal RNA polymerase.

We're also interested in transcription regulation in Bacteria (with Tracy Nixon, Penn State; and with Haw Yang, Princeton). The latter efforts are devoted to understanding a family of molecular motors that are involved in bacterial transcription activation, the enhancer-binding proteins (EBP) of the NtrC family (NtrC, NtrC1, NtrC4). We have made substantial progress in understanding the structural basis of the regulated assembly and function of NtrC (EM DataBase entry 1218) and gained insight about the interaction of EBP with the sigma (σ54) factor.

• De Carlo S., Lin S.-C., Taatjes D.J., and A. Hoenger (2010). Molecular Basis of Transcription Initiation in Archaea.
• J. Batchelor, M. Doucleff, C. Lee, K. Matsubara, S. De Carlo, C. J. Heideker, M. Lamers, J. G. Pelton, D. E. Wemmer (2008). Structure and regulatory mechanism of A. aeolicus NtrC4: variability and evolution in bacterial transcriptional regulation. J. Mol. Biology 384, 1058-1075.
• De Carlo S., Boisset N., and A. Hoenger (2008). High-resolution Single-particles 3-D Analysis on GroEL prepared by Cryo-negative Staining. Micron 39, 934-943.
• Chen B., Doucleff M., Wemmer D., De Carlo S., Huang H., Nogales E., Hoover T., Kondrashkina E., Guo L., and B. T. Nixon (2007). ATP Ground- and Transition States of the Enhancer Binding AAA+ ATPases Support Complex Formation with their Target Protein,σ54. Structure 15, 429-440. COVER
• Kostek S., Grob P., De Carlo S., Lipscomb S., Garczarek F., and E. Nogales (2006). Molecular Architecture and Conformational Flexibility of human RNA Polymerase II. Structure 14, 1691-1700.
• De Carlo S., Chen B., Hoover T., Kondrashkina E., Nogales E., and B. T. Nixon (2006). The structural basis of regulated assembly and function of the transcriptional activator NtrC. Genes & Dev. 20, 1485-1495. COVER
• Jawhari A., Uhring M., De Carlo S., Crucifix C., Tocchini-Valentini G., Moras D., Schultz P. and Poterszman A. (2006). Structure and oligomeric state of human transcription factor TFIIE. EMBO Rep. 7, 500-505.
• De Carlo S., Fiaux H., and A Marca-Martinet C. (2004). Electron Cryo-Microscopy Reveals the Mechanism of Action of Propranolol on Artificial Membranes. J. Lip. Res. 14, 61-76.
• De Carlo S., Carles C., Riva M. and Schultz P. (2003). Cryo-negative staining reveals conformational flexibility within yeast RNA Polymerase I. J. Mol. Biol. 329, 891-902.
• De Carlo S., El-Bez C., Alvarez-Rúa C., Borge J. and Dubochet J. (2002). Cryo-negative staining reduces electron beam sensitivity of vitrified biological particles. J. Struct. Biol. 138, 216-226. COVER

• De Carlo S. Cryo-electron microscopy: plunge freezing. In: Handbook of Cryopreparation Methods for Electron Microscopy. Ed. CRC Press (2008). ISBN: 9780849372278.
• De Carlo S. Cryo-negative staining. In: Handbook of Cryopreparation Methods for Electron Microscopy. Ed. CRC Press (2008). ISBN: 9780849372278.