%0 Journal Article %1 claw %A Harjes, E %A Kitamura, A %A Zhao, W %A Morais, M C %A Jardine, P J %A Grimes, S %A Matsuo, H %D 2012 %J Nucleic Acids Res %K phage %N 19 %P 9953-9963 %R 10.1093/nar/gks724 %T Structure of the RNA claw of the DNA packaging motor of bacteriophage Φ29 %U http://www.ncbi.nlm.nih.gov/pubmed/22879380 %V 40 %X Bacteriophage DNA packaging motors translocate their genomic DNA into viral heads, compacting it to near-crystalline density. The Bacillus subtilis phage 29 has a unique ring of RNA (pRNA) that is an essential component of its motor, serving as a scaffold for the packaging ATPase. Previously, deletion of a three-base bulge (18-CCA-20) in the pRNA A-helix was shown to abolish packaging activity. Here, we solved the structure of this crucial bulge by nuclear magnetic resonance (NMR) using a 27mer RNA fragment containing the bulge (27b). The bulge actually involves five nucleotides (17-UCCA-20 and A100), as U17 and A100 are not base paired as predicted. Mutational analysis showed these newly identified bulge residues are important for DNA packaging. The bulge introduces a 33-35° bend in the helical axis, and inter-helical motion around this bend appears to be restricted. A model of the functional 120b pRNA was generated using a 27b NMR structure and the crystal structure of the 66b prohead-binding domain. Fitting this model into a cryo-EM map generated a pentameric pRNA structure; five helices projecting from the pRNA ring resemble an RNA claw. Biochemical analysis suggested that this shape is important for coordinated motor action required for DNA translocation.

@article{claw, abstract = {Bacteriophage DNA packaging motors translocate their genomic DNA into viral heads, compacting it to near-crystalline density. The Bacillus subtilis phage 29 has a unique ring of RNA (pRNA) that is an essential component of its motor, serving as a scaffold for the packaging ATPase. Previously, deletion of a three-base bulge (18-CCA-20) in the pRNA A-helix was shown to abolish packaging activity. Here, we solved the structure of this crucial bulge by nuclear magnetic resonance (NMR) using a 27mer RNA fragment containing the bulge (27b). The bulge actually involves five nucleotides (17-UCCA-20 and A100), as U17 and A100 are not base paired as predicted. Mutational analysis showed these newly identified bulge residues are important for DNA packaging. The bulge introduces a 33-35° bend in the helical axis, and inter-helical motion around this bend appears to be restricted. A model of the functional 120b pRNA was generated using a 27b NMR structure and the crystal structure of the 66b prohead-binding domain. Fitting this model into a cryo-EM map generated a pentameric pRNA structure; five helices projecting from the pRNA ring resemble an RNA claw. Biochemical analysis suggested that this shape is important for coordinated motor action required for DNA translocation.}, added-at = {2014-02-23T13:10:47.000+0100}, author = {Harjes, E and Kitamura, A and Zhao, W and Morais, M C and Jardine, P J and Grimes, S and Matsuo, H}, biburl = {https://www.bibsonomy.org/bibtex/2196cff92f3cd5711a6352cbeb2a63426/ross_mck}, description = {Structure of the RNA claw of the DNA packa... [Nucleic Acids Res. 2012] - PubMed - NCBI}, doi = {10.1093/nar/gks724}, interhash = {0e3ee5197fa0cc5bc6a05e506c7f0ad3}, intrahash = {196cff92f3cd5711a6352cbeb2a63426}, journal = {Nucleic Acids Res}, keywords = {phage}, month = oct, number = 19, pages = {9953-9963}, pmid = {22879380}, timestamp = {2014-02-23T13:10:47.000+0100}, title = {Structure of the RNA claw of the DNA packaging motor of bacteriophage Φ29}, url = {http://www.ncbi.nlm.nih.gov/pubmed/22879380}, volume = 40, year = 2012 }