SARAH MARIE AUCLAIR, PH.D.
50 William Ave
Meriden, CT 06451
Phone (508) 736-5548
Wesleyan University, Middletown, Connecticut May 2010
Doctor of Philosophy in Molecular Biology and Biochemistry
Wheaton College, Norton, Massachusetts May 2003
Bachelor of Arts in Biology; Minor in Chemistry
Associate Research Scientist
Nanobiology Institute, Department of Cell Biology, Yale University, CT
Investigated the ability of large 25 nm nanolipo-particles (nanodiscs) to work in our standard fusion assay and the ability for these discs to allow the fusion pore to expand. Optimized the protocol to prepare small 45 nm vesicles containing membrane proteins using a DNA origami ring as a size template. Tested various tethers (DNA, PNA, Click chemistry) to be used as a way to bring two vesicles together to improve fusion.
Achievements: Completed work on two research projects, one of which is currently being written up in manuscript form. This work has been presented in 3 research seminars.
Feb 2013-April 2014
Nanobiology Institute, Department of Cell Biology, Yale University, CT
Examined the role of the VAMP2-TMD in vesicle fusion by using TMD mutants in a T-SNARE SUVs and VAMP2-nanodisc content release assay. Developed new content release methods to measure vesicle fusion that would allow the use of Synaptotagmin and physiological lipids. Established a purification method for Synaptotagmin to help stabilize the protein and reduce the degradation products over time.
Achievements: Completed work on three research projects and presented the research in 3 seminars. This work resulted in 3 manuscripts.
Jun 2010-Dec 2012
Department of Pharmaceutical Sciences, University of Connecticut, CT
Developed a protocol for creating SPase I-nanodiscs. Examined the solvent accessibility of Δ2-75 SPase I tryptophan residues in the presence and absence of lipid vesicles and signal peptide. Investigated the relationship between the open and closed states of SecA when interacting with various ligands. Used analytical ultracentrifugation and alanine scanning mutagenesis to identify critical residues in the formation and stabilization of the SecA dimer.
Achievements: Wrote an American Heart Association grant, a L’Oreal women in science grant, and 2 NIH NRSA grants. Published a first author article in the peer-reviewed journal of Protein Sciences and another paper in Biochemistry.
Sept 2003-May 2010
Graduate Research Assistant
Molecular Biology and Biochemistry Department, Wesleyan University, CT
Developed a novel method using FRET to map the signal peptide binding site on Escherichia coli SecA. Employed FRET to determine the orientation of solution state dimeric SecA based on the 5 different X-ray structures of dimeric SecA published to date.
Achievements: Published two first author articles in the peer-reviewed journal of Biochemistry. Awarded the Peterson Fellowship for graduate study in biochemistry.
January & Summer 2003
Protein Chemistry Department, Critical Therapeutics, Inc., MA
Assisted with initial laboratory set up and purchasing of equipment at this biotech startup. Worked on development of a method to purify anti-peptide antibodies on a peptide affinity column and screened using an anti-peptide ELISA with poly-D-lysine-coated plates. Sera, flow-through, and elution fractions were tested for endotoxins using the Limulus amebocyte lysate assay.
Biology Department, Wheaton College, MA
Researched the genetic diversity of spotted salamanders and diamondback terrapins by analyzing genetic polymorphisms. Recreated medieval medicines from Bauds Leechbook for various ailments and tested their effectiveness of inhibition for common bacterial strains.
Achievements: Results of the medieval medicines project were published in Anglo-Saxon England. Awarded 4 Wheaton Foundation grants to support my research. Awarded the Julia R. Lange Fellowship to help support my first year of graduate school.
Research and Development, Wyeth Research, MA
Musculoskeletal Sciences Discovery Research Internship Program
Cloned NF-ĸB-activating kinase (NAK) and TANK into yeast two-hybrid vectors. Set up yeast two-hybrid system with controls and screened two human libraries using full-length NAK as the bait.
Sanitary Microbiology Lab, RI State Department of Health Laboratories, RI
General lab maintenance to ensure all equipment was functioning properly. Kept detailed records on incubator and fridge temperatures for quality control. Prepared media for general lab use.
Achievements: Awarded a Wheaton Fellowship to support this internship.
Microbiology and Molecular Genetics Department, University of Vermont, VT
• Soluble and membrane protein purification using both gel filtration, ion exchange, and affinity columns with gravity and FLPC.
• Transmembrane peptide purification on HPLC
• Proteoliposomes –SUVs, LUVs, and IMVs
• MSP and NLP nanodiscs
• DNA origami rings to template SUV formation of a particular size
• Click chemistry, DNA, and PNA tethers to link two liposomes together
• Fluorescence techniques-Protein labeling, FRET, anisotropy, TIRF, acrylamide and 10-DN quenching.
• Microscale Thermophoresis (MST) binding assays
• Dynamic Light Scattering (DLS)
• Circular Dichroism (CD) Spectroscopy
• UV-IR spectroscopy
• Content release and lipid mixing assays, ATPase assays
• Chemical and UV crosslinking or proteins to identify interactions
• SDS-PAGE, Native-PAGE, Western blots
• Molecular cloning, Site-directed mutagenesis, RAPD-PCR, DNA electrophoresis, DNA extraction from tissues and bacteria
• Other skills: analytical ultracentrifugation, transmission electron microscopy, Yeast two-hybrid, Kirby-Bauer inhibition test, ELISA, Endotoxin testing, Antibody purification, and Cell culture.
• Trained two postdocs, four graduate students and four undergraduates in general laboratory techniques and in fluorescence spectroscopy.
• Ordered lab supplies and equipment.
• Requested quotes for large equipment purchases.
• Serviced and maintained equipment.
• Managed and trained 15 student computer help desk consultants.
• Scheduled shifts.
• Responsible for the safety and security of the campus computer center and its equipment.
• Assisted with performance evaluations and decisions regarding promotions.
BUSINESS AND MANAGERIAL EXPERIENCE
Vitae Essentia (previously Ladybug Soapworks), Meriden, CT
Designed and developed a small business that manufactures handmade natural soap for retail and wholesale. Grew the company to be financially solvent after the first year. Main responsibilities include supply purchasing, accounting, manufacture and packaging of product, marketing, and design and maintenance of the company website. Developed recipes and designs for over 40 products.
HAS Computer Lab, Wesleyan University, CT
Managed and trained 12 student employees and scheduled shifts. Provided general help with logging into computers, printing, and basic application problems. Acted as a telephone dispatcher for classroom support to assist faculty or dispatch a technician to fix problems in the “smart” classrooms.
Help Desk Student Manager
Kollett Academic Computing Center, Wheaton College, MA
Responsible for managing the center and training student employees. Provided assistance to students in the computing center and throughout the campus with computer related issues. This included configuring new computers to assess the campus network and resolving software and printing issues.
Wu, Z. Bello, O., Auclair, S.M., Vennekate, W., Krishnakumar, S.S., and Karatekin, E. (in preparation) SNARE proteins dilate fusion pores through crowding.
Wu, Z., Auclair, S.M., Bello, O., Vennekate, W., Dudzinski, N., Krishnakumar, S.S., Karatekin, E. (2016) Nature Scientific Reports. Nanodisc-cell fusion: control of fusion pore nucleation and lifetimes by SNARE protein transmembrane domains.
Bello, O., Auclair, S.M., Krishnakumar, S.S., and Rothman, J.E. (2016) Langmuir. Using ApoE Nanolipoprotein Particles To Analyze SNARE-Induced Fusion Pores.
Wang, J., Bello, O., Auclair, S.M., Wang, J., Coleman, J., Pincet, F., Krishnakumar, S.S., Sindelar, C.V., and Rothman, J.E. (2014) PNAS. Calcium sensitive ring-like oligomers formed by Synaptotagmin.
Wowor, A.J., Yan, Y., Auclair, S.M., Yu, D., Zhang, J., May, E.R., Gross, M.L., Kendall, D.A., and Cole, J. (2014) Biochemistry. Analysis of SecA Dimerization in Solution.
Auclair, S.M., Mukerji, I., and Oliver, D.B. (2013) Biochemistry. Identification of the dimer orientation of solution state Escherichia coli SecA using Förster resonance energy transfer.
Auclair, S.M., Bhanu, M.K., and Kendall, D.A. (2011) Protein Science. Signal peptidase I: cleaving the way to mature proteins. (Highlighted in the front of the journal.)
Auclair, S.M., Moses, J., Musial-Siwek, M., Kendall, D.A., Oliver, D.B., and Mukerji, I. (2010) Biochemistry. Mapping of the signal peptide-binding domain of Escherichia coli SecA using Förster resonance energy transfer.
Peterson Fellowship (Wesleyan University) 2009
Julia R. Lange Fellowship (Wheaton College) 2003
Wheaton Foundation Grant (Wheaton College) 2001, 2002
Wheaton Fellowship (Wheaton College) 2001, 2002
Nellie S. Smith Scholarship (Wheaton College) 2000-2003
Newport Country Club Scholarship 1999
Amy Booth Meyers Memorial Scholarship 1999
Montgomery Education Association Scholarship 1999
AD HOC REVIEWER
Cell Stress and Chaperones 2012-Present
TEACHING AND TEACHING ASSISTANT EXPERIENCES
Invited guest speaker for a Careers in Health and Biomedical Sciences Seminar March 2010
St. Joseph’s College, CT
Taught a freshman career seminar class on a career path in the biological sciences.
MB&B337/537 Molecular Basis of Pathogenicity Spring 2005
Wesleyan University, CT
Assisted the professor with class material preparation and exam corrections, and held office hours for students.
MB&B103 The Green Gene Revolution or GMOs in the Garden Spring 2004 Wesleyan University, CT
Taught one full lecture for the professor and acted as a teaching assistant for the course lectures including correcting exams and holding office hours.
MB&B205 Principles of Biology I: Cell Biology and Molecular Basis of Heredity Fall 03, Fall 04 Wesleyan University, CT
Acted as a teaching assistant for the course lectures including correcting exams and holding office hours.
MB&B215 Principles of Biology I: Laboratory Fall 03, Fall 04
Wesleyan University, CT
Independently taught one laboratory course section per week including material and lecture preparation, running and teaching a laboratory section, correcting laboratory reports, and holding office hours.
Nanobiology Research Talks, Yale University, CT 5 Seminars between 2013-2015
Invited Seminar; Yale University, CT January 23, 2013
Biochemistry Research Seminar; Wesleyan University, CT. March 24, 2010
Invited Seminar; University of Connecticut Health Center, CT. March 17, 2010
Invited Seminar; Yale University, CT. March 3, 2010
Invited Seminar; Boston Biomedical Research Institute, MA. January 28, 2010
MB&B Research Seminar; Wesleyan University, CT. 8 Seminars in 2003-2009
PUBLISHED ABSTRACTS AND PRESENTATIONS
Over 25 poster presentations of my work at various conferences.
Beta Beta Beta National Biological Society 2003-Present
Biophysical Society 2007-Present
American Heart Association 2012-2013
Society for Economic Botany 2011-2012
Society of Ethnobiology 2010-2011
American Society of Microbiology 2006-2007
Graduate Student Association Social Committee, Wesleyan University 2006-2007
Graduate Student Association Benefits Committee, Wesleyan University 2005-2007
Graduate Student Association Housing Committee, Wesleyan University 2004-2007
Graduate Student Association Student Judiciary Board Member, Wesleyan University 2004-2005
Hope House President, Wheaton College 2001-2002