Controlling bacteriophage phi29 DNA-packaging motor by addition or discharge of a peptide at N-terminus of connector protein that interacts with pRNA.
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.
Sun
Jianhe
J
Department of Pathobiology, Purdue Cancer Center and Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA.
Cai
Ying
Y
Moll
Wulf-Dieter
WD
Guo
Peixuan
P
D017103:Bacillus Phages / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D002213:Capsid / Q000648:ultrastructure; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D042003:DNA Packaging; D019281:Dimerization; D015870:Gene Expression; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*; D014771:Virion / Q000502:physiology; D019065:Virus Assembly
D036022:Capsid Proteins; D010455:Peptides; D012367:RNA, Viral; C097975:portal protein, bacteriophage phi29