American Journal of Microbiological Research
ISSN (Print): 2328-4129 ISSN (Online): 2328-4137 Website: http://www.sciepub.com/journal/ajmr Editor-in-chief: Apply for this position
Open Access
Journal Browser
Go
American Journal of Microbiological Research. 2014, 2(1), 1-7
DOI: 10.12691/ajmr-2-1-1
Open AccessArticle

Stem Mutants in the N-terminal Domain of the Phage P22 Tailspike Protein

Chris Palmer1, Jeremie Williams1, Dexter Dean2, 3, Sam Johnson1, Hongzhuan Wu1, Boakai K. Robertson1, Doba Jackson2 and Robert Villafane1,

1Program in Microbiology, Alabama State University, Montgomery, AL, USA

2Department of Chemistry and Biochemistry, Huntingdon College, Montgomery, AL, USA

3Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS, USA

Pub. Date: December 26, 2013

Cite this paper:
Chris Palmer, Jeremie Williams, Dexter Dean, Sam Johnson, Hongzhuan Wu, Boakai K. Robertson, Doba Jackson and Robert Villafane. Stem Mutants in the N-terminal Domain of the Phage P22 Tailspike Protein. American Journal of Microbiological Research. 2014; 2(1):1-7. doi: 10.12691/ajmr-2-1-1

Abstract

The P22 tailspike protein is an intensely studied protein whose structure and sequence has been described. However, a study, describing important protein interactions related to its function at the N-terminal domain, has been lacking. The P22 tailspike protein (TSP) consists of three identical polypeptide chains of 666aa. The first 108 of the 666aa in the P22 TSP form a trimeric N-terminal domain (NTD). Each of the three chains of the trimeric NTD contributes to the formation of a dome-like structure. Our studies suggest that a short stretch of amino acids located within the first fifteen amino acids of the P22 TSP NTD is critical for the stability of the dome structure formed by the first 108aa of the P22 TSP NTD. The first 23aa are located within this dome-like structure and have been dubbed the “stem” of the NTD. Although amino acid residues in the first 15aa (lower stem) are critical, deletion analysis and in vitro assembly studies implicate the rest of the stem in additional stabilizing interactions. Our studies implicate a common protein-protein interaction motif made up of interchain hydrophobic contacts between adjacent chains

Keywords:
protein-protein interactions Salmonella tailspike protein phage assembly

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Figures

Figure of 4

References:

[1]  Campbell, A., “Comparative molecular biology of lambdoid phages,” Annual Review of Microbiology, 48,193-222, 1994.
 
[2]  Casjens, S.R., Thuman-Commike, P.A., “Evolution of mosaically related tailed bacteriophage genomes seen through the lens of phage P 22 virion assembly,” Virology, 411, 393-415, 2012.
 
[3]  Botstein, D., Waddell, C. H., King J., “Mechanism of head assembly and DNA encapsulation in Salmonella phage P 22. I. Genes, proteins, structures and DNA metabolism,” Journal of Molecular Biology, 80, 669-695, 1973.
 
[4]  King, J., Lenk, E. V., Botstein, D., “Mechanism of head assembly and DNA encapsulation in Salmonella phage P 22. II. Morphogenetic pathway,” Journal of Molecular Biology, 80, 697-731, 1973.
 
[5]  Hartwieg, E., Bazinet, C., King, J., “DNA injection apparatus of phage P22,” Biophysical Journal, 49, 24-26, 1984.
 
[6]  Chiang, J., Weigele, P., King, J., Chiu, W., Jiang, W., “Cryo-EM asymmetric reconstruction of bacteriophage P 22 reveals organization of its DNA packaging and infecting machinery,” Structure, 14, 1073-1082, 2006.
 
[7]  Lander, G. C., Khayat, R., L,i R., Prevelige, P.E., Potter, C. S., Carragher, B., Johnson, J. E., “The P 22 tail machine at subnanometer resolution reveals the architecture of an infection conduit,” Structure, 17, 789-798, 2009.
 
[8]  Lander, G.C., Tang, L., Casjens, S.R., Gilcrease, E. B., Prevelige, P., Poliakov, A., Potter, C. S., Carraher, B., Johnson, J. E., “The structure of an infectious P 22 virion shows the signal for headful DNA packaging,” Science, 312, 1791-1795, 2006.
 
[9]  Steinbacher, S., Miller, S., Baxa, U., Budisa, N., Weintraub, A., Seckler, R., Huber, R., “Phage P 22 tailspike protein: Crystal structure of the head-binding domain at 2.3 Å, fully refined structure of the endorhamnosidase at 1.56 Å resolution, and the molecular basis of O-antigen recognition and cleavage,” Journal of Molecular Biology, 267, 865-880, 1997.
 
[10]  Steinbacher, S., Seckler, R., Miller, S., Streipe, B., Huber, R., Reinemer, P., “Crystal structure of the P 22 tailspike protein: Interdigitated subunits in a thermostable trimmers,” Science, 265, 383-386, 1994.
 
[11]  The PyMOL Molecular Graphics System, Version 1.5.0.4 Schrödinger, LLC.
 
[12]  Israel, J. V., Anderson, T. F, Levine, M., “In vitro morphogenesis of phage P 22 from phage heads and base-plate parts,” Proceedings of the National Academy of Sciences, 57, 284-291, 1967.
 
[13]  Schwarz, J. J., Berget, P. B., “The isolation and sequence of missense and nonsense mutations in the cloned bacteriophage P 22 TSP gene,” Genetics, 121, 635-649, 1989.
 
[14]  Villafane, R., Costa, S., Ahmed, R., Salgado, C., “Conservation of the N-terminus of some phage tails proteins,” Archives of Virology, 150, 2609-2621, 2005.
 
[15]  Robinson, A. S., King, J., “Disulfide-bonded intermediate on the folding and assembly pathway of a non-disulfide bonded protein,” Nature Structural Biology, 4, 450-455, 1997.
 
[16]  Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning: A Laboratory Manual, 2nd edition, CSH Press, New York, 1989.
 
[17]  Villafane, R., “Construction of phage mutants,” Methods in Molecular Biology, 501, 223-237, 2009.
 
[18]  Villafane, R., Fleming, A., Haase-Pettingell, C., “Isolation of suppressors of temperature-sensitive folding mutants,” Journal of Bacteriology, 176:137-142, 1994.
 
[19]  Berget, P. B., Poteete, A. R., Sauer, R. T., “Control of phage P 22 tail protein expression by transcription termination,” Journal of Molecular Biology, 164, 561-572, 1983.
 
[20]  Kamei, D.T., Liu, C., Haase-Pettingell, C., King, J. A., Wang, D. I. C., Blankschtein, D., “Understanding viral partitioning in two-phase nonionic micellar systems: Role of attractive interactions between viruses and micelles,” Biotechnology and Bioengineering, 78, 190-202, 2002.
 
[21]  Haase-Pettingell, C., Betts, S., Raso, S.W., Stuart, L., Robinson, A., King, J., “Role of cysteine residues in the in vivo folding and assembly of the phage P 22 tailspike,” Protein Science, 10, 397-410, 2001.
 
[22]  Weigele, P. R., Haase-Pettingell, C., Campbell, P. G., Gossard, D.C., King, J. “Stalled folding mutants in the triple beta -helix domain of the phage P 22 tailspike adhesion,” Journal of Molecular Biology, 354, 1103-1117, 2005.
 
[23]  Copeland, R.A. Methods for Protein Analysis: A Practical Guide To Laboratory Protocols, Chapman and Hall Press, New York, 1994, pp. 59-98.
 
[24]  Roskams, J., Rodgers, L., Electrophoretic separation of proteins and nucleic acids. In: Roskams J, Rodgers L (eds), Lab Ref: A handbook of recipes, reagents, and other reference tools for use at the bench. Cold Spring Harbor Press, New York, 2002, pp. 63-91.
 
[25]  Chandler, D., “Interfaces and the driving force for hydrophobic assembly,” Nature, 437, 640-647, 2005.
 
[26]  Pace, C. N., Fu, H., Fryar, K. L., Landua, J., Trevino, S.R., Shirley, B.A., Hendricks, M.M., Iimura, S., Gajiwa, K., Scholtz, J. M., Grimsley, G. R., “Contribution of hydrophobic interaction to protein stability,” Journal of Molecular Biology, 408, 514-528, 2011.
 
[27]  Saha, R. P., Bahardur, R. P., Chakrabarti, P., “Interresidue contacts in proteins and protein-protein interfaces and their use in characterizing the homodimeric interface,” Journal of Proteome Research, 4, 1600-1609, 2005.
 
[28]  Tsai, C-J., Lin, S. L., Wolfson, H.J., Nussinov, R., “Studies on protein-protein interfaces: A statistical analysis of the hydrophobic effect,” Protein Science, 6, 53-64, 1997.
 
[29]  Moreira, I. S., Fernandes, P.A., Ramos, M. J., “Hot spots–A review of the protein-protein interface determinant amino acid residues,” Proteins, 68, 803-812, 2007.
 
[30]  Glaser, F., Steinberg, D.M., Vakser, I. A., Ben-Tal, N., “Residue frequencies and pairing preferences at protein-protein interfaces,” Proteins, 43, 89-102, 2001.
 
[31]  Keskin, O., Gursoy, A., Ma, B., Nussinov, R., “Principles of protein-protein interactions: What are the preferred ways for proteins to interact?” Chemical Reviews, 108, 1225-1244, 2008.
 
[32]  Dill, K. A., “Dominant forces in protein folding,” Biochemistry, 29, 7133-7155, 1990.
 
[33]  Maurides, P.A., Schwarz, J. J., Berget, P. B., “Intragenic suppression of a capsid-assembly defective P 22 tailspike mutation”, Genetics, 125, 673-681, 1989.