Nucleic Acids Research Advance Access originally published online on October 4, 2006
Nucleic Acids Research 2006 34(19):5482-5490; doi:10.1093/nar/gkl701
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Nucleic Acids Research, 2006, Vol. 34, No. 19 5482-5490
© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Controlling bacteriophage phi29 DNA-packaging motor by addition or discharge of a peptide at N-terminus of connector protein that interacts with pRNA
1 Department of Pathobiology, Purdue Cancer Center and Weldon School of Biomedical Engineering, Purdue University West Lafayette, IN 47907, USA 2 School of Agriculture and Biology, Shanghai Jiaotong University Shanghai, 201101, People's Republic of China
*To whom correspondence should be addressed at Purdue Cancer Center, B-36 Hansen Life Science Research Building, Purdue University, West Lafayette, IN 47907, USA. Tel: +1 765 494 7561; Fax: +1 765 496 1795; Email: guop{at}purdue.edu
Received July 12, 2006. Revised September 1, 2006. Accepted September 6, 2006.
Bacteriophage phi29 utilizes a motor to translocate genomic DNA into a preformed procapsid. The motor contains six pRNAs, an enzyme and one 12-subunit connector with a central channel for DNA transportation. A 20-residue peptide containing a His-tag was fused to the N-terminus of the connector protein gp10. This fusion neither interfered with procapsid assembly nor affected the morphology of the prolate-shaped procapsid. However, the pRNA binding and virion assembly activity were greatly reduced. Such decreased functions can be switched back on by the removal of the tag via protease cleavage, supporting the previous finding that the N-terminus of gp10 is essential for the pRNA binding. The DNA-packaging efficiency with dimeric pRNA was more seriously affected by the extension than with monomeric pRNA. It is speculated that the fusion of the tag generated physical hindrance to pRNA binding, with greater influence for the dimers than the monomers due to their size. These results reveal a potential to turn off and turn on the motor by attaching or removing, respectively, a component to outer part of the motor, and offers an approach for the inhibition of viral replication by using a drug or a small peptide targeted to motor components.
Present address: Wulf-Dieter Moll, Department for Agrobiotechnology, IFA-Tulln, University of Natural Resources and Applied Life Sciences, Vienna, Austria