Associate Professor, Center for RNA Molecular Biology (Faculty since 1996)
Education: Ph.D.: Department of Biochemistry, University of Alabama at Birmingham
Post-doc: Department of Biology, Indiana University
Secondary Appointment in: Department of Biochemistry
Principal Investigator and Director of the Cell and Molecular Biology Training Grant/Program
Office Location: Wood Blgd. W106
Office Phone: 216-368-4779
Office FAX: 216-368-2010
Laboratory Location: W103
Laboratory Phone: 216-368-0381
RNase P is a ribonucleoprotein enzyme with a catalytic RNA subunit that depends on a small protein cofactor for molecular recognition. Thus, it serves as a facile model system to explore fundamental aspects of molecular recognition, RNA-protein interactions and RNA-metal ion interactions. We are pursuing these lines of investigation using the standard physical and spectroscopic tools as well as developing new approaches including collaborative efforts at application of kinetic isotope effects (with Vernon Anderson) and Raman spectroscopy (with Paul Carey).
Two of the fundamental questions for both RNA and protein enzymes that catalyze phosphoryl transfer are “What is the mechanism of the reaction?” and “How do active site functional groups influence the chemical bonding of the substrate to achieve catalysis?” Both the chemical mechanism and active site interactions influence the bonding environment of the atoms undergoing reaction, which can be assessed by measuring the effects of isotopic substitution at these positions. An important outgrowth of our collaborative studies of ribozyme catalysis has been the development of methods to determine isotope effects for enzymatic and non-enzymatic phosphoryl transfer reactions involving RNA. These studies promise to provide important new information on the mechanism of RNA catalyzed reactions and tests of current hypotheses concerning the role of specific active site interactions.
Pubmed Link to Publications
Google Scholar Link to Publications
Zahler, N.H. and Harris, M.E. (2004). “Site-Specific Modification of RNA” ” in Handbook of RNA Biochemistry, Wiley. Hartmann, Binderief, Shon and Westhof, eds.
Harris, M.E. and Christian, E.L. (2004). “Probing RNA Structure by Photoaffinity Crosslinking With 4-Thiourine and 6-Thioguanosine” in Handbook of RNA Biochemistry, Wiley. Hartmann, Binderief, Shon and Westhof, eds.
Zahler, N.H., Sun, L., Christian, E.L. and Harris, M.E. (2005). The pre-tRNA nucleotide base and 2'-hydroxyl at N(-1) contribute to fidelity in tRNA processing by RNase P. J. Molec. Biol. 345(5):969-85.
Anderson, V.E., Harris ME and Cassano AG (2006) “Nucleophile Isotope Effects” in Isotope Effects in Chemistry and Biology. CRC Press. Kohen A, Limbach H-H (eds)
Anderson, V.E., Ruszczysky, M. and Harris, M.E. (2006). Nucleophilic activation in enzyme catalysis. Chem. Rev. 106(8):3236-51.
Guo X, Campbell FE, Christian EL, Anderson VE and Harris ME. (2006) RNA-dependent folding and stabilization of C5 protein during assembly of the E. coli RNase P holoenzyme. J. Molec. Biol. 360(1):190-203
Christian EL, Johnson, K, Perrera, N and Harris, ME (2006). Evidence for a role for the P4 metal binding site in enhancing catalytic metal ion affinity by substrate positioning. RNA 12(8):1463-7.
Sun L, Campbell FE and Harris ME (2006) Substrate specific effects of C5 protein lead to uniformity in binding and catalysis by RNase P. EMBO J. 25(17):3998-4007
Harris ME and Anderson VE (2007) “Chemistry of Natural Ribozymes”. Encyclopedia of Chemical Biology. Wiley.
Sun L and Harris ME (2007). Binding of C5 protein to the P RNA ribozyme increases catalytic rate by influencing active site metal ion affinity. RNA 34:1-11.
Cassano AG, Wang B, Anderson DR, Previs S, Harris ME, Anderson VE. (2007). Inaccuracies in selected ion monitoring determination of isotope ratios obviated by profile acquisition: nucleotide 18O/16O measurements. Anal. Biochem. 367(1):28-39.
Dai, Q, Frederiksen, J, Anderson VE, Harris ME, and Piccirilli, J (2008) Efficient synthesis of 2'-[18O]-uridine and its incorporation into oligonucleotides: A new tool for mechanistic studies of phosphoryl transfer reactions by isotope effect analyses. J. Org. Chem. 73(1):309-11
Harris, ME and Christian EL (2008) Understanding the Role of Metal Ions in RNA Folding and Function: Lessons from RNase P, a Ribonucleoprotein Enzyme. In Biophysics of Non-protein Encoding RNAs. Springer, Inc.
Harris ME, Cassano AG. (2008). Experimental analyses of the chemical dynamics of ribozyme catalysis. Curr Opin Chem Biol. 12(6):626-39.
Schroeder LA, Gries TJ, Saecker RM, Record MT Jr, Harris ME, DeHaseth PL. (2008). Evidence for a tyrosine-adenine stacking interaction and for a short-lived open intermediate subsequent to initial binding of Escherichia coli RNA polymerase to promoter DNA. J Mol Biol. 385(2):339-49.
Harris ME and Christian EL (2009). RNA Crosslinking Methods. in Methods in Enzymology, Vol. 468, Biophysical, Chemical and Functional Probes of RNA Structure, Interactions and Folding, Part A., Herschlag, D, (ed.), Academic Press, pp.127-146
Christian EL, Anderson VE, Carey PC and Harris ME. (2010). A Raman spectroscopic signature for RNA metal ion interactions in solution. Biochemistry. 49(13):2869-79.PMID: 20180599.
Koutmou KS, Zahler NH, Kurz JC, Campbell FE, Harris ME, Fierke CA. (2010). Protein-precursor tRNA contact leads to sequence-specific recognition of 5' leaders by bacterial ribonuclease P. J Mol Biol. 396(1):195-208. PMID: 19932118
Sun L, Campbell FE, Yandek LE, Harris ME. (2010). Binding of C5 protein to P RNA enhances the rate constant for catalysis for P RNA processing of pre-tRNAs lacking a consensus G(+1)/C(+ 72) pair. J Mol Biol. 395(5):1019-37. PMID: 19917291
Harris ME and Yandeck LE. (2010). “Challenges in substrate recognition by RNase P: Facing up to the biological context” in Protein Reviews Volume 10: Ribonuclease P. pp 135-152. Altman S and Lui F, eds. Springer New York. ISBN 978-1-4419-1141-4
Christian EL, Anderson VE, Carey PR, Harris ME. (2010). A quantitative Raman spectroscopic signal for metal-phosphodiester interactions in solution. Biochemistry. 49(13):2869-79.PMID: 20180599
Harris ME, Dai Q, Gu H, Kellerman DL, Piccirilli JA, Anderson VE. (2010). Kinetic isotope effects for RNA cleavage by 2'-O- transphosphorylation: Nucleophilic activation by specific base. J Am Chem Soc. 132(33):11613-21.
Christian EL, Anderson VE, Harris ME. (2011). Deconvolution of Raman spectroscopic signals for electrostatic, H-bonding, and inner-sphere interactions between ions and dimethyl phosphate in solution. J Inorg Biochem. In press