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CURRICULUM VITAE Saddef Haq, Ph.D. Program in Toxicology University of Maryland Baltimore School of Medicine

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CURRICULUM VITAE Saddef Haq, Ph.D. Program in Toxicology University of Maryland Baltimore School of Medicine Beyond the Dinoflagellate Transcriptome: Validation of Protein Production via Biochemical Analysis and Mass Spectrometry Item Type dissertation Authors Haq, Saddef Publication Date 2018 Abstract Dinoflagellates are members of the Alveolata (meaning “with cavities”), a monophyletic group of single cell protists which includes apicomplexans and ciliates that exhibit a diverse mode of nutrition, ranging from predation to photo autotrophy to int... Keywords bioluminescence; dinoflagellates; toxin synthesis; Acetyl-CoA Carboxylase; Dinoflagellida; Polyketide Synthases; Proteomics Download date 27/09/2021 16:23:13 Link to Item http://hdl.handle.net/10713/8970 CURRICULUM VITAE Saddef Haq, Ph.D. Program in Toxicology University of Maryland Baltimore School of Medicine Institute of Marine and Environmental Technology 701 E. Pratt Street Baltimore, Maryland 21202 Email: [email protected] EDUCATION 2013-2018 University of Maryland, Baltimore, MD Program in Toxicology Doctor of Philosophy 2006-2008 Rutgers University, New Brunswick, NJ Bachelor of Science in Biochemistry 2003-2006 Union County College, Cranford, NJ Associates of Science in Biology EMPLOYMENT HISTORY July 2013-Present Graduate Research Assistant Graduate Program in Life Sciences, Program in Toxicology University of Maryland, Baltimore, Baltimore, MD Lab of Dr. Allen R. Place at the Institute of Marine and Environmental Technology, Baltimore, MD. Sep 2010-May 2013 Senior Research Technician Center for Infection and Immunity Columbia University, School of Public Health New York, NY Mar 2009-Dec 2009 Laboratory Technician Unigene Laboratories Inc. Boonton, NJ Oct 2008-Feb 2009 Sampling Laboratory Technician Imclone Systems Inc. Branchburg, NJ HONORS AND AWARDS June 2016-June 2017 Ratcliffe Environmental Entrepreneurship Fellowship Stipend, tuition, and technology support Additional award to start company and develop product prototype November 2015 Travel Award for US Harmful Algal Bloom Symposium PUBLICATIONS Haq S., Oyler B.L., Williams E.P., Khan M.M., Goodlett D.R., Nita-Lazar A., Bachvaroff T.R., Place A.R. Investigating a Multi-Modular Polyketide Synthase in Amphidinium carterae. 2018. In Preparation. Haq, S.; Bachvaroff, T. R.; Place, A. R. Characterization of Acetyl-CoA Carboxylases in the Basal Dinoflagellate Amphidinium carterae. Mar. Drugs 2017, 1–10. Haq, S.; Bachvaroff, T. R.; Place, A. R. Phylogenetic Analysis of Acetyl CoA Carboxylases in Dinoflagellates. In 17th ICHA Proceedings; 2017, 138-141. Rodriguez, J. D.; Haq, S.; Bachvaroff, T.; Nowak, K. F.; Nowak, S. J.; Morgan, D.; Cherny, V. V.; Sapp, M. M.; Bernstein, S.; Bolt, A.; Decoursey, T. E.; Place, A. R.; Smith, S. M. E. Identification of a Vacuolar Proton Channel that Triggers the Bioluminescent Flash in Dinoflagellates. PLoS One 2017, 12, 1–24. Cai, P.; He, S.; Zhou, C.; Place, A. R.; Haq, S.; Ding, L.; Chen, H.; Jiang, Y.; Guo, C.; Xu, Y.; Zhang, J.; Yan, X. Two new karlotoxins found in Karlodinium veneficum (strain GM2) from the East China Sea. Harmful Algae 2016. Bhat, N.; Tokarz, R.; Jain, K.; Haq, S.; Weatherholtz, R.; Chandran, A.; Karron, R.; Reid, R.; Santosham, M.; O’Brien, K. L.; Lipkin, W. I. A Prospective Study of Agents Associated with Acute Respiratory Infection among Young American Indian Children. Pediatr. Infect. Dis. J. 2013. Tokarz, R.; Haq, S.; Sameroff, S.; Howie, S. R. C.; Lipkin, W. I. Genomic analysis of coxsackieviruses A1, A19, A22, enteroviruses 113 and 104: Viruses representing two clades with distinct tropism within enterovirus c. J. Gen. Virol. 2013, 94, 1995–2004. Tokarz, R.; Hirschberg, D. L.; Sameroff, S.; Haq, S.; Luna, G.; Bennett, A. J.; Silva, M.; Leguia, M.; Kasper, M.; Bausch, D. G.; Lipkin, W. I. Genomic analysis of two novel human enterovirus C genotypes found in respiratory samples from Peru. J. Gen. Virol. 2013, 94, 120–127. Tokarz, R.; Briese, T.; Morris, G.; Ideh, R.; Chimah, O.; Ebruke, B.; Desai, A.; Haq, S.; Sameroff, S.; Howie, S. R. C.; Lipkin, W. I. Serotype analysis of streptococcus pneumoniae in lung and nasopharyngeal aspirates from children in the Gambia by Masstag PCR. J. Clin. Microbiol. 2013, 51, 995–997. Tokarz, R.; Firth, C.; Madhi, S. A.; Howie, S. R. C.; Wu, W.; Sall, A. A.; Haq, S.; Briese, T.; Ian Lipkin, W. Worldwide emergence of multiple clades of enterovirus 68. J. Gen. Virol. 2012, 93, 1952–1958. PRESENTATIONS Haq S, Bachvaroff TR, Oyler B, Place AR. Acetyl-CoA Carboxylases in Dinoflagellates: Fueling the Polyketide Synthase Pathways. US Symposium on Harmful Algae. Baltimore, MD. November 11-17, 2017. Speed Talk and Poster Presentation. Haq S, Bachvaroff TR, Oyler B, Place AR. Acetyl-CoA Carboxylases in Dinoflagellates: Fueling the Polyketide Synthase Pathways. Gordon Research Conference: Mycotoxins and Phycotoxins. Easton, MA. June 18-23, 2017. Poster Presentation. Haq S, Bachvaroff TR, Oyler B, Place AR. Acetyl-CoA Carboxylases in Dinoflagellates: Fueling the Polyketide Synthase Pathways. Gordon Research Seminar: Mycotoxins and Phycotoxins. Easton, MA. June 17-18, 2017. Oral Presentation. Haq S, Bachvaroff TR, Goodlett DR, Place AR. Acetyl-CoA Carboxylases in Dinoflagellates: Fueling the Polyketide Synthase Pathways. International Conference on Harmful Algae. Florianopolis, Brazil. October 9-14, 2016. Poster Presentation. Haq S, Bachvaroff TR, Goodlett DR, Place AR. Acetyl-CoA Carboxylases in Dinoflagellates: Fueling the Polyketide Synthase Pathways. International Marine Biotechnology Conference. Baltimore, MD. August 29-September 2, 2016. Oral Presentation. Haq S, Bachvaroff TR, Goodlett DR, Place AR. Acetyl-CoA Carboxylases in Dinoflagellates: Fueling the Polyketide Synthase Pathways. Oceans and Human Health Grantee Meeting. Research Triangle Park, NC. April 13-14, 2016. Poster Presentation. Haq S, Bachvaroff TR, Williams E, Goodlett DR, Place AR. One Pathway-Two Products: A Polyketide Synthase in Dinoflagellates Results in Both Toxin and Fatty Acid Production. Mass Spectrometry School in Biotechnology and Medicine. Dubrovnik, Croatia. July 5-11, 2015. Poster Presentation. Haq S, Bachvaroff TR, Williams E, Goodlett DR, Place AR. One Pathway-Two Products: A Polyketide Synthase in Dinoflagellates Results in Both Toxin and Fatty Acid Production. US Symposium on Harmful Algae. Long Beach, CA. November 15-19, 2015. Speed Talk and Poster Presentation. Abstract Title: Beyond the Dinoflagellate Transcriptome: Validation of Protein Production via Biochemical Analysis and Mass Spectrometry Saddef Haq, Doctor of Philosophy, 2018 Dissertation Directed By: Allen R. Place, PhD, Professor University of Maryland, Center for Environmental Science Institute of Marine and Environmental Technology Dinoflagellates are members of the Alveolata (meaning “with cavities”), a monophyletic group of single cell protists which includes apicomplexans and ciliates that exhibit a diverse mode of nutrition, ranging from predation to photo autotrophy to intracellular parasitism. Dinoflagellates are both primary producers and consumers in the food web, sometimes at the same time, and best known for their dominant role in causing harmful algal blooms. In keeping with their unusual nuclear chromatin architecture (condensed throughout the cell cycle) the degree to which dinoflagellates use transcriptional responses to alter mRNA expression levels appears limited. In addition, it appears dinoflagellates have extensive gene duplications for most biological processes. For these reasons, the goal for this dissertation was to measure protein levels directly for important cellular phenotypes. We began with characterizing a proton selective channel (Hv1) that was hypothesized to be involved in bioluminescence in Lingulodinium polyedrum with goals of elucidating function, location, and production of this protein. Next, we focused on acetyl CoA carboxylases (ACC) which are responsible for the rate limiting step in fatty acid (FAS) and polyketide synthases (PKS). We find the presence of these proteins in thirteen dinoflagellates surveyed and show there is distribution in the plastid and cytosol. In Amphidinium carterae we verify protein production with high performance liquid chromatography-coupled to tandem mass spectrometry (LC-MS/MS), otherwise known as shotgun proteomics as well as compare transcript and protein levels across a diel cycle. Lastly, we focused on a multi-modular polyketide protein in A. carterae with implications in toxin and fatty acid production due to the domain arrangement which is indicative of a noniterative trans-AT PKS process. We show successful inhibition of 14C acetate incorporation into toxins and fatty acids with the addition of the ketosynthase specific (KS) fatty acid inhibitor cerulenin. Partial protein production is verified by western blotting and LC-MS/MS analysis of KS peptides reveals a strong covalent bond with cerulenin addition. Taken together the work discussed in this thesis has resulted in a better understanding of three different cellular processes in dinoflagellates using a shotgun proteomic approach. Future work continuing this trend of protein characterization in dinoflagellates will help elucidate many uncharacterized pathways in dinoflagellate biology. Beyond the Dinoflagellate Transcriptome: Validation of Protein Production via Biochemical Analysis and Mass Spectrometry by Saddef Haq Dissertation submitted to the Faculty of the Graduate School of the University of Maryland, Baltimore in partial fulfillment of the requirements for the degree of Doctor of Philosophy 2018 ©Copyright 2018 Saddef Haq All Rights Reserved DEDICATION
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