Dr. Fred W. Perrino
Professor of Biochemistry
B.S., Ohio State University, 1979
Ph.D. (Biological Sciences), University of Cincinnati, 1986
Telephone: (336) 716-4349
Electronic mail: fperrino@wfubmc.edu
Enzymes that replicate, repair, and disassemble the human genome
3’à5’ DNA exonucleases
During the multi-step processes of DNA replication,
repair, recombination, and apoptosis DNA is remodeled by enzymes containing 3’à5’ exonuclease activities. The major 3’à5’ exonuclease activity in human cells is catalyzed by the enzyme encoded by the TREX1 gene that was discovered in the Perrino laboratory. While the existence of exonucleases in human cells has been recognized for nearly forty years, only recently have the genes encoding these enzymes, including TREX1 and the closely related TREX2, been identified. Thus, there is currently insufficient information about the 3’à5’ exonucleases in human cells to understand the mechanisms and the molecular pathways in which these proteins function.
The Perrino laboratory is collaborating with the Hollis laboratory to examine structure, mechanism, and function of the TREX enzymes. The goal of these experiments is to gain a better understanding of the biochemistry of the TREX 3’à5’ exonucleases.
TREX1 mutations have been identified in families and in individuals with Aicardi-Goutières Syndrome (AGS), Familial Chilblain Lupus (FCL), and in some individuals with Systemic Lupus Erythematosus (SLE), Sjögren´s syndrome and Raynaud’s phenomenon. The identification of TREX1 mutations in these related autoimmune disorders and the linkage of TREX1 to a cell death pathway in collaborative work with the Lieberman laboratory provide compelling evidence for a TREX1 role in the DNA disassembly process during apoptosis in order to prevent autoimmune response to DNA.
Our studies are directed at unraveling the mechanistic details of the TREX1 enzyme to better understand the molecular actions of this DNA processing enzyme and the pathways through which mutations in this enzyme lead to human autoimmune disease.
DNA polymerization of damaged DNA
The human genome is susceptible to damage by many chemicals produced naturally in cells or by environmental exposure. The biological consequence of this damage is relevant to mutagenesis, to carcinogenesis, and to cancer therapeutics. Despite our recognition of DNA damage as a path to mutagenesis our knowledge concerning the response of the human DNA polymerases to the damage inflicted on DNA is limited. The Perrino laboratory is collaborating with the Hollis laboratory in a structure-based mechanistic analysis of the human Y-family DNA polymerases and adducted DNA to understand this process.
Recent publications:
Lee-Kirsch, MA, Gong, M, Chowdhury, D, Senenko, L, Engel, K, Lee YA, de Silva, U, Bailey, SL, Witte, T, Vyse, TJ, Kere, J, Pfeiffer, C, Harvey, S, Wong, A, Koskenmies, S, Hummel, O, Rohde, K, Schmidt, RE, Dominiczak, AF, Gahr, M, Hollis, T, Perrino, FW, Lieberman, J and Huber, N (2007) Mutations in the 3’à5’ DNA exonuclease TREX1 are associated with systemic lupus erythematosus, Nature Genetics, 39, 1065-1067.
Lee-Kirsch, MA, Chowdhury, D, Harvey, S, Gong, M, Senenko, L, Engel, K, Pfeiffer, C, Hollis, T, Gahr, M, Perrino, FW, Lieberman, J and Huber, N (2007) A mutation in TREX1 that impairs susceptibility to granzyme A-mediated cell death underlies familial chilblain lupus, J. Mol. Med., 85, 531-537.
Rice, G, Newman, WG, Dean, J, Patrick, T, Parma, R, Flintoff, K, Robins, P, Harvey, S, Hollis, T, O’Hara, A, Herrick, AL, Bowden, AP, Bonthron, DT, Perrino, FW, Lindahl, T, Barnes, DE and Crow, YJ (2007) Heterozygous mutations in TREX1 cause familial chilblain lupus and dominant Aicardi-Goutières syndrome, Amer. J. Human Genetics, 80, 811-815.
de Silva, U, Choudhury, S, Bailey, SL, Perrino, FW and Hollis T (2007) The crystal structure of TREX1 explains the 3' nucleotide specificity and reveals a polyproline II helix for protein partnering, J. Biol. Chem., 282, 10537-10543.
Harrigan, JA, Fan, J, Momand, J, Perrino, FW, Bohr, VA, and Wilson III, DA (2007) WRN exonuclease activity on DNA 3’ blocking termini, and comparison to APE1, TREX1, and p53, Mechanisms of Ageing and Development, 128, 259-266.
Upton, D.C., Wang, X., Blans, P., Perrino, F.W., Fishbein, J.C., and Akman, S.A. (2006) Mutagenesis by exocyclic alkylamino purine adducts in Escherichia coli, Mutation Research, 599:1-10.
Upton, D.C., Wang, X., Blans, P., Perrino, F.W., Fishbein, J.C., and Akman, S.A. (2006) Replication of N2-ethyldeoxyguanosine DNA adducts in human 293 cells, Chem. Res. Toxicol., 19:960-967.
Chowdhury, D, Beresford, PJ, Zhu, P, Zhang, D, Sung, J-S, Demple, B, Perrino, FW, Lieberman, J (2006) The exonuclease TREX1 is in the SET complex and acts in concert with NM23-H1 to degrade DNA during granzyme A-mediated cell death, Molecular Cell, 23:133-142.
Kirby, TW, Harvey, S, DeRose, EF, Chalov, S, Perrino, FW, Schaaper, R, London, RE and Pedersen, L (2006) Structure of the interspecies complex formed by the E. coli proofreading exonuclease domain of DNA polymerase III and the P1 Phage-encoded homolog of q, HOT, J. Biol. Chem., 281, 38466-38471.
Perrino, F. W., Harvey, S., Blans, P., Gelhaus, S.L., LaCourse, W. R., Fishbein, J.C. (2005) Polymerization Past the N2-Isopropylguanine and the N6-Isopropyladenine DNA Lesions with the Translesion Synthesis DNA Polymerases η and i and the Replicative DNA Polymerase α, Chem. Res. Toxicol., 18:1451-61.
Perrino, FW, Harvey, S, McMillin, S, Hollis, T (2005) The human TREX2 3'à5' exonuclease structure suggests a mechanism for efficient nonprocessive DNA catalysis, J. Biol. Chem., 280, 15212-15218.
Perrino, FW, Krol, A, Harvey, S, Xu, J, Zheng, S, Horita, DA, Hollis, T, Meyers, DA, Isaacs, WB (2004) Sequence Variants in the 3’à5’ Deoxyribonuclease TREX2: Identification in a Genetic Screen and Effects On Catalysis by the Recombinant Proteins, Advances in Enzyme Regulation, 44, 37-49.