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
- 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) - 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) - 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) - 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) - 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) - 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 - 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)
- 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)
- 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. - 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
- 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
- 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)
- 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. - 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. - 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. - 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. - 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. - 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. - Chen, C. and P. Guo. 1997. Sequential action of six
DNA-packaging pRNA during phage phi29 genomic DNA translocation. J.
Virology 71:3864-3871. - 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. - 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. - Trottier, M. and P. Guo. 1997. Approaches to Determine
Stoichiometry of Viral Assembly Components. J. Virology, 71:487-494.
- 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. - 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. - 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. - Lee, C. S. and P. Guo. 1995. Sequential interactions of
structural proteins in phage phi29 procapsid assembly. J Virol 69:5024-5032.
- 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. - 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. - 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. - 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. - 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, - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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. - 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
- 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. - 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. - 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. - Guo, P. 1994. Editor’s Introduction: Principles,
perspectives, and potential applications in viral assembly. Seminars in
Virology 5(1):1-3. - Guo, P. 1988 Mechanism of DNA packaging in ds-DNA
bacteriophages. Virogica Sinica 2:229-2