Structure of protein from neck of bacteriophage infecting Staphylococcus aureus

Barbora Břenková1, Marta Šiborová1, Pavol Bárdy2, Roman Pantůček2,
Pavel Plevka1

1Central European Institute of Technology, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic;

2Department of Experimental Biology, Faculty of Science, Masaryk University, Kotlářská 267/2, 611 37 Brno, Czech Republic; correspondence should be addressed to pavel.plevka@ceitec.muni.cz

 

Bacteriophage phi812 is a lytic phage from the family Myoviridae. Phi812 infects most staphylococcal strains including those resistant to antibiotics [1]. The structure of this bacteriophage was previously determined by cryo electron microscopy [2] but functions of individual proteins remain unclear. Here we present our work towards resolving the structure of protein gp99 that forms the neck region of phi812. The protein may play a role in a regulation of the genome release from the virion during infection. Gp99 was cloned and expressed in E. coli and purified by affinity and size exclusion chromatography. A suitable crystallization condition was found and X-ray diffraction data with a resolution of 2.3 A were collected. To solve the phase problem, a variety of methods had been tried, including molecular replacement, heavy atom soaking and co-crystallization, and seleno-methionine incorporation. The structure of gp99 will help explain the mechanism of bacterial infection by bacteriophage. As there is a rising number of antibiotic resistant bacterial strains causing severe illnesses, phage therapy has a high potential. For this purpose, detailed knowledge of bacteria-phage interactions is vital.

 

1.         Pantůček, R., Rosypalová, A., Doškař, J., Kailerová, J., Růžičková, V., Borecká, P., Snopková, S., Horváth, R., Götz, F., Rosypal, S. “The Polyvalent Staphylococcal Phage Phi 812: Its Host-Range Mutants and Related Phages.” Virology., U.S. National Library of Medicine, 5 July 1998, www.ncbi.nlm.nih.gov/pubmed/9657943.

 

2.         Nováček, J., Šiborová, M., Benešík, M., Pantůček, R., Doškař, J., Plevka, P. “Structure and Genome Release of Twort-like Myoviridae Phage with a Double-Layered Baseplate.” Proceedings of the National Academy of Sciences of the United States of America., U.S. National Library of Medicine, 16 Aug. 2016, www.ncbi.nlm.nih.gov/pubmed/27469164.