Peixuan Guo

Information:

Professor of Molecular Virology 

Purdue Cancer Research Center

Purdue University

Hansen B036
201 South University Street

West Lafayette, IN 47907-2064

Phone # (765) 494-7561

Fax # (765) 496-1795

E-mail:[email protected]

Research Interest:

We found that a 120-base RNA (pRNA) was required for DNA packaging (Guo, et al. Science 236: 690-694, 1987). Our recent findings imply that phi29 DNA packaging is accomplished by a mechanism similar to the driving of a bolt with a hex nut (J. Virol. 71:3864,1997), and six pRNAs form a hexagonal complex to gear the DNA translocating machine (J.Viro 71:487,1997; Mol.Cell 2:147,1998; Cell 94:147,1998). The current focus is to find out how phi29 pRNA performs a task carried out by protein enzymes.

RNA enzymes, called ribozymes, can be used for treatment and prevention of diseases in plants, humans, and animals. Although it is clear that hammerhead ribozymes could cleave specific RNA in the test tube, the efficiency of cleavage in the cells of animals and plants is dramatically reduced due to misfolding or degradation of the ribozymes in cells by exonucleases. We have developed a novel vector for hammerhead ribozyme to ensure appropriate folding of the ribozyme and to protect ribozymes from degradation by exonucleases

The capsid of dsDNA bacterial viruses contains a six-fold symmetrical DNA-translocating machine or connector embedded in a protein shell with a five-fold rotational symmetry.  We proposed a model to show that the relative motion of the two symmetrical mismatched rings could provide a driving force for DNA transportation.  Analogous to the six cylinders in the engine of a car, sequential action is a way to turn the motor.  The finding that six copies of pRNA were bound to the connector and worked sequentially is potentially relevant to this model.  To make two rings rotate relatively, at least one additional component is needed to provide a propelling force. We found that the packaging of phi29 DNA requires ATP as energy source to provide the driving force for DNA translocation.   Interaction of RNA with ATP has never been reported before. We also found that RNA is part of the ATPase.  Alternative construction and realization of pRNA will provide a physical force for DNA translocation. This project is to study how ATP can cause conformational change of pRNA. 

Our goal is to develop an in vivo gene delivery system for cancer therapy based on the in vitro assembly system of bacteriophage ø29.  Methods to package the gene of interest and the assembly of the delivery particles have been accomplished in this lab as following:
 1. A highly efficient in vitro viral packaging system for the encapsidation of specific and non-specific DNA has been developed.
 2. Dual plasmid vectors with the expression of reporter genes in both prokaryotic and eukaryotic cells have been constructed.
 3. We are able to assemble infectious ø29 virion in vitro with all proteins produced from cloned genes and both DNA and RNA synthesized in vitro.
 4. A new strategy for complete intracellular immunization to viral infection has been developed.
 5. We are working on the construction of delivery particles carrying specific ligand(s) to recognize receptor(s) on the surface of human cells for specific targeted delivery. 

Representative Publications:

Original Papers in Referred Journals

  1. Huang L. and Guo P. 2003. Use of PEG and Acetone to Attain Highly Active and Soluble DNA Packaging Protein Gp16 of Phi29 for ATPase Assay. Virology (in press)
  2. Huang L and Guo P. 2003.  Use of PEG to acquire highly soluble DNA-packaging enzyme gp16 of bacterial virus phi29 for stoichiometry quantification.  J. Virol Meth.  (in press) 
  3. Hoeprich S, Qi G, Guo S, Shu D, Wang Y and Guo P. 2003. Phi29 pRNA as a Hammerhead Ribozyme Escort to Destroy Hepatitis B Virus, Gene Therapy - Nature Publishers (in press)
  4. Shu D and Guo P. 2002.  Only one pRNA hexamer but multiple copies of the DNA-packaging protein gp16 are needed for the motor to package bacterial virus phi29 genomic DNA Virology (in press)
  5. Shu D, Huang L, Hoeprich S and Guo P.  Construction of phi29 DNA-packaging RNA (pRNA) Monomers, Dimers and Trimers With Variable Sizes and Shapes as Potential Parts for Nano-devices.  Journal of Nanoscience and Nanotechnology (in press)
  6. Shu D and Guo P.A viral RNA that binds ATP and contains a motif similar to an ATP-binding aptamer from SELEX. J Biol Chem. 2002 Nov 20  http://www.jbc.org/cgi/reprint/M209895200v1
  7. Hoeprich S, Guo P. Computer Modeling of Three-dimensional Structure of DNA-packaging RNA (pRNA) Monomer, Dimer, and Hexamer of Phi29 DNA Packaging Motor. J Biol Chem. 2002 Jun 7;277(23):20794-803 (http://www.jbc.org/cgi/content/full/277/23/20794#F2)
  8. Mat-Arip Y, Garver K, Chen C, Sheng S, Shao Z, Guo P. 2001.Three-dimensional Interaction of Phi29 pRNA Dimer Probed by Chemical Modification Interference, Cryo-FM,and Cross-linking. J Biol Chem. 276(35):32575-84. (http://www.jbc.org/cgi/reprint/276/35/32575.pdf)
  9. Zhang C. Trottier, M. and Guo, P.  2001 Chemical modification patterns of  active and inactive as well as  procasid-bound and unbound DNA packaging RNA of bacterial virus phi29. Virology, 281:281-293.
  10. M.Trottier, Y. Mat-Arp, C. Zhang, C. Chen, S. Sheng, Z. Shao, and P. Guo. 2000. Probing the structure of monomers and dimers of the bacterial virus phi29 hexamer RNA complex by chemical modification. RNA 6(9): 1257-1266. http://www.journals.cup.org/ows-bin/article?spii=S1355838200992501
  11. C. Chen, S. Sheng., Z. Shao, P. Guo. 2000. A Dimer as a Building Block in Assembling RNA. J. Biol. Chem.275(23): 17510-17516. http://www.jbc.org/cgi/content/full/275/23/17510
  12. K. Garver and P. Guo. 2000. Mapping the inter-pRNA interaction of phage phi29 pRNA by site-specific photoaffinity crosslinking. J. Biol. Chem. 275(4): 2817-2824 .(http://www.jbc.org/cgi/content/full/275/4/2817?maxtoshow=&HITS=10&hits=10&RE)
  13. Mohammad, T., C.  Chen,  P. Guo and H. Morrison. 1999. Photo-induced cross-linking of RNA by cis-ph(phen)2Cl2+ and cis-ph(phen)(phi)l2+. Bioorganic and Medicinal Chemistry Letters 9:1703-1708.
  14. Aggarwa, N.,  H. HogenEsch, P. Guo, A. North, M. Suckow and S. Mittal. 1999. Biodegradable Alginate Microspheres as a Delivery System for Naked DNA. Can J. Vet. Res. 63:148-152.
  15. Chen, C., Zhang C. and P. Guo. 1999.  Sequence requirement for hand-in-hand interaction to form hexameric RNA complex for phage phi29 DNA packaging. RNA 5:805-818.
  16. Guo, P., C. Zhang, Chen, C., K. Garver, M. Trottier and  C. Chen. 1998. Inter-RNA Interaction of phage phi29 pRNA to Form a Hexameric Complex for DNA Transportation. Mol. Cell 2:149-155.
  17. Trottier, M., Garver, K., Zhang, C. and  P. Guo. 1997. DNA-packaging pRNA as target for complete inhibition of viral assembly  in vitro  and in vivo. Nucleic Acids Symposium Series. 36:187-189.
  18. Chen, C., Trottier, M. C. and P. Guo. 1997.  New approaches to  stoichiometry determination and mechanism investigation on RNA involved in intermediate reactions. Nucleic Acid Symp Series. 36:190-193.
  19. Chen, C. and P. Guo. 1997.  Sequential action of six DNA-packaging pRNA during phage phi29 genomic DNA translocation. J. Virology 71:3864-3871.
  20. Garver, K. and P. Guo. 1997. Boundary of pRNA functional domains and minimum pRNA sequence requirement for specific connector binding and DNA packaging of phage phi29.  RNA 3:1068-1079.
  21. Chen, C. and P. Guo. 1997.  Magnesium-induced conformational change of pRNA for procapsid recognition and binding during bacteriophage phi29 DNA packaging.  J. Virology 71: 495-500.
  22. Trottier, M. and P. Guo.  1997. Approaches to Determine Stoichiometry of Viral Assembly Components.  J. Virology, 71:487-494.
  23. Zhang, C. L., T. Tellinghuiesn  and P. Guo.  1997. Use of circular permutation to assess six bulges and four loops of DNA-packaging pRNA of bacteriophage phi29. RNA 3:315-322.
  24. Huang, Q., Y. Mat-Arip, and P. Guo. 1997.  Sequencing of a 5.5-kb DNA fragment and identification of a gene coding for a subunit of the helicase/primase complex of avian Laryngotracheitis virus (ILTV). Virus Gene 15:(2): 119-121.
  25. Trottier, M., C. L. Zhang, and P. Guo.  1996. Complete inhibition of virion assembly in vivo  with mutant pRNA essential for phage phi29 DNA packaging. J Virol 70:55-61.
  26. Lee, C. S. and P. Guo.  1995. Sequential interactions of structural proteins in phage phi29 procapsid assembly. J Virol 69:5024-5032.
  27. Lee, C. S. and P. Guo.  1995. In vitro assembly of infectious virions of ds-DNA phage phi29 from cloned gene products and synthetic nucleic acids. J Virol 69:5018-5023.
  28. Zhang, C. L., T. Tellinghuiesn, and P. Guo.  1995. Confirmation of the helical structure of the 5’/3’ termini of the essential DNA packaging pRNA of phage phi29. RNA 1:1041-1050.
  29. Zhang, C. L., M. Trottier, and P. Guo.  1995. Circularly permuted viral pRNA active and specific in the packaging of bacteriophage phi29 DNA. Virology 207:442-451.
  30. Zhang, C. L., K. Garver, and P. Guo.  1995. Inhibition of phage phi29 assembly by antisense oligonucleotides targeting viral pRNA essential for DNA packaging. Virology 211:568-576.
  31. Scholz, E. and P. Guo. 1995. Construction of Recombinant Avian Infectious Laryngotracheitis Virus with TK Gene disrupted by ß-gal Coding Sequence. In Imm Viral Inf. Proc. 3rd Intl Cong Vet. Virol, 379-384,
  32. Guo, P., E. Scholz, B. Maloney, and E. Welniak.  1994. Construction of recombinant avian infectious laryngotracheitis virus expressing the b-galactosidase gene and DNA sequencing of the insertion region. Virology 202:771-781.
  33. Lee, C. and P. Guo.  1994. A highly sensitive system for the assay of in vitro  viral assembly of bacteriophage phi29  of Bacillus subtilis. Virology 202:1039-1042.
  34. Scholz, E., R. E. Porter, and P. Guo.  1994. Differential diagnosis of infectious laryngotracheitis from other avian respiratory disease by a simplified PCR procedure. J Virol Meth.  50:313-322.
  35. Zhang, C. . L., C. -S. Lee, and P. Guo.  1994. The proximate 5’ and 3’ ends of the 120-base viral RNA (pRNA) are crucial for the packaging of bacteriophage phi29 DNA. Virology 201:77-85.
  36. Guo, P., E. Scholz, J. Turek, R. Nordgren, and B. Maloney.  1993. Assembly pathway of avian infectious laryngotracheitis virus. Am J Vet Res 54:2031-2039.
  37. Scholz, E., E. Welniak, T. Nyholm, and P. Guo.  1993. An avian hepatoma cell line for the cultivation of infectious laryngotracheitis virus and for the expression of foreign genes with a mammalian promotor. J Virol Meth 43:273-286.
  38. Scholz, E., C. L. Zhang, and P. Guo.  1993. Transactivation of the early SV40 promoter by avian infectious laryngotracheitis virus in avian hepatoma cells. J Virol Meth 45:291-301.
  39. Lee, C. -S. and P. Guo.  1993. Tracking and elimination of a interfering polypeptide co-expressed with the vaccinia virus mRNA Capping Enzyme. Protein Expression and Purification 4:114-119.
  40. Guo, P., S. Erickson, W. Xu, N. Olson, T. S. Baker, and D. Anderson.  1991. Regulation of the phage phi29 prohead shape and size by the portal vertex. Virology 183:366-373.
  41. Guo, P., B. Rajogopal, D. Anderson, S. Erickson, and C. -S. Lee.  1991. sRNA of bacteriophage phi29 of B.subtilis mediates DNA packaging of phi29 proheads assembled in E. coli. Virology 185:395-400.
  42. Guo, P. and B. Moss.  1990. Interaction and mutual stabilization of the two subunits of vaccinia virus mRNA capping enzyme co-expressed in Escherichia coli. Proc Natl Acad Sci USA 87:4023-4027.
  43. Guo, P., S. Goebel, M. Perkus, J. Taylor, E. Norton, G. Allen, B. Languet, P. Desmettre, and E. Paoletti.  1990. Coexpression by vaccinia virus recombinants of equine herpesvirus 1 glycoproteins gp13 and 14 results in potentiated immunity. J Virol 64:2399-2406.
  44. Guo, P. 1990. Characterization of the gene and an antigenic determinant of equine herpesvirus 1 glycoprotein 14 with homology to gB-equivalent glycoprotein of other herpesvirus. Gene 87:249-255.
  45. Guo, P., S. Goebel, S. Davis, M. Perkus, B. Languet, P. Desmettre, G. Allen, and E. Paoletti.  1989. Expression of the equine herpesvirus type 1 gene encoding glycoprotein gp13 in recombinant vaccinia virus and protection of immunized animals. J Virol 63:4189-4198.
  46. Guo, P.  1988. Inhibition of novobiocin, coumermycin A, oxolinic acid, nalidixic, ethidium bromide and ATP analogue on the packaging of phi29 DNA in vitro. Virologica Sinica 2:198-205.
  47. Guo, P., C. Peterson, and D. Anderson.  1987. Prohead and DNA-gp3-dependent ATPase activity of the DNA packaging protein gp16 of bacteriophage phi29. J Mol Biol 197:229-236.
  48. Guo, P., S. Erickson, and D. Anderson.  1987. A small viral RNA is required for in vitro packaging of bacteriophage phi29 DNA. Science 236:690-694.
  49. Guo, P., S. Bailey, J. W. Bodley, and D. Anderson.  1987. Characterization of the small RNA of the bacteriophage phi29 DNA packaging machine. Nucleic Acids Res. 15:7081-7090.
  50. Guo, P., C. Peterson, and D. Anderson.  1987. Initiation events in in vitro packaging of bacteriophage phi29 DNA-gp3. J Mol Biol 197:219-228.
  51. Guo, P., S. Grimes, and D. Anderson.  1986. A defined system for in vitro packaging of DNA-gp3 of the Bacillus subtilis bacteriophage phi29. Proc Natl Acad Sci USA 83:3505-3509

Reviews

  1. Guo P.  2002.  Structure and function of phi29 hexameric RNA that drives the viral DNA packaging motor.  Progress in Nucleic Acids Research 72: 415-474.
  2. Guo P.  2002. Methods for Structural and Functional Analysis of pRNA in Bacterial Virus Phi29  DNA Packaging Motor. Acta Biochim Biophys Sin.    34(5):533-543.
  3. Guo, P. and M. Trottier.  1994. Biological and biochemical properties of the small viral RNA (pRNA) essential for the packaging of the double-stranded DNA of phage phi29. Seminars in Virology 5:27-37.
  4. Guo, P.  1994. Editor’s Introduction: Principles, perspectives, and potential applications in viral assembly. Seminars in Virology 5(1):1-3.
  5. Guo, P. 1988 Mechanism of DNA packaging in ds-DNA bacteriophages. Virogica Sinica 2:229-2