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Structure and Assembly of TP901-1 Virion Unveiled by Mutagenesis.

Stockdale SR, Collins B, Spinelli S, Douillard FP, Mahony J, Cambillau C, van Sinderen D - PLoS ONE (2015)

Bottom Line: Bacteriophages of the Siphoviridae family represent the most abundant viral morphology in the biosphere, yet many molecular aspects of their virion structure, assembly and associated functions remain to be unveiled.Fourteen mutations located within the structural module of TP901-1 were created; twelve mutations were designed to prevent full length translation of putative proteins by non-sense mutations, while two additional mutations caused aberrant protein production.Based on the information obtained, we propose a functional and assembly model of the TP901-1 Siphoviridae virion.

View Article: PubMed Central - PubMed

Affiliation: School of Microbiology, University College Cork, Western Road, Cork, Ireland.

ABSTRACT
Bacteriophages of the Siphoviridae family represent the most abundant viral morphology in the biosphere, yet many molecular aspects of their virion structure, assembly and associated functions remain to be unveiled. In this study, we present a comprehensive mutational and molecular analysis of the temperate Lactococcus lactis-infecting phage TP901-1. Fourteen mutations located within the structural module of TP901-1 were created; twelve mutations were designed to prevent full length translation of putative proteins by non-sense mutations, while two additional mutations caused aberrant protein production. Electron microscopy and Western blot analysis of mutant virion preparations, as well as in vitro assembly of phage mutant combinations, revealed the essential nature of many of the corresponding gene products and provided information on their biological function(s). Based on the information obtained, we propose a functional and assembly model of the TP901-1 Siphoviridae virion.

No MeSH data available.


Related in: MedlinePlus

Schematic representation of the 22 annotated genes of the structural module of phage TP901-1.Mutagenesis of individual genes of the TP901-1 structural module showed gene products to be essential (+) or non-essential (-) for creating infectious phage virions. The efficiency of plaquing is displayed for mutants of non-essential gene products. The genomic regions encoding capsid and tail morphogenesis proteins are indicated and such assignments are based on data described in the text.
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pone.0131676.g001: Schematic representation of the 22 annotated genes of the structural module of phage TP901-1.Mutagenesis of individual genes of the TP901-1 structural module showed gene products to be essential (+) or non-essential (-) for creating infectious phage virions. The efficiency of plaquing is displayed for mutants of non-essential gene products. The genomic regions encoding capsid and tail morphogenesis proteins are indicated and such assignments are based on data described in the text.

Mentions: The structural module of TP901-1 is represented by a polycistronic operon, approximately 20 kb in length, transcribed from a promoter located upstream of the gene encoding the predicted small terminase subunit [7, 8, 12]. This structural module of TP901-1 is predicted to encode 22 proteins (Fig 1) [7]. Functional analyses have previously been performed for a number of TP901-1 structural proteins (and by inference those of the closely related phage Tuc2009) [6, 9, 11, 13, 16, 19, 56, 80, 81]. Due to the conserved nature of phage structural proteins, functions of several TP901-1 proteins can be predicted based on a number of well characterized phage prototypes [42, 55, 82–84]. However, some TP901-1 proteins do not exhibit significant similarity to known proteins and thus their functional assignment is based on literature and bioinformatic analyses.


Structure and Assembly of TP901-1 Virion Unveiled by Mutagenesis.

Stockdale SR, Collins B, Spinelli S, Douillard FP, Mahony J, Cambillau C, van Sinderen D - PLoS ONE (2015)

Schematic representation of the 22 annotated genes of the structural module of phage TP901-1.Mutagenesis of individual genes of the TP901-1 structural module showed gene products to be essential (+) or non-essential (-) for creating infectious phage virions. The efficiency of plaquing is displayed for mutants of non-essential gene products. The genomic regions encoding capsid and tail morphogenesis proteins are indicated and such assignments are based on data described in the text.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493119&req=5

pone.0131676.g001: Schematic representation of the 22 annotated genes of the structural module of phage TP901-1.Mutagenesis of individual genes of the TP901-1 structural module showed gene products to be essential (+) or non-essential (-) for creating infectious phage virions. The efficiency of plaquing is displayed for mutants of non-essential gene products. The genomic regions encoding capsid and tail morphogenesis proteins are indicated and such assignments are based on data described in the text.
Mentions: The structural module of TP901-1 is represented by a polycistronic operon, approximately 20 kb in length, transcribed from a promoter located upstream of the gene encoding the predicted small terminase subunit [7, 8, 12]. This structural module of TP901-1 is predicted to encode 22 proteins (Fig 1) [7]. Functional analyses have previously been performed for a number of TP901-1 structural proteins (and by inference those of the closely related phage Tuc2009) [6, 9, 11, 13, 16, 19, 56, 80, 81]. Due to the conserved nature of phage structural proteins, functions of several TP901-1 proteins can be predicted based on a number of well characterized phage prototypes [42, 55, 82–84]. However, some TP901-1 proteins do not exhibit significant similarity to known proteins and thus their functional assignment is based on literature and bioinformatic analyses.

Bottom Line: Bacteriophages of the Siphoviridae family represent the most abundant viral morphology in the biosphere, yet many molecular aspects of their virion structure, assembly and associated functions remain to be unveiled.Fourteen mutations located within the structural module of TP901-1 were created; twelve mutations were designed to prevent full length translation of putative proteins by non-sense mutations, while two additional mutations caused aberrant protein production.Based on the information obtained, we propose a functional and assembly model of the TP901-1 Siphoviridae virion.

View Article: PubMed Central - PubMed

Affiliation: School of Microbiology, University College Cork, Western Road, Cork, Ireland.

ABSTRACT
Bacteriophages of the Siphoviridae family represent the most abundant viral morphology in the biosphere, yet many molecular aspects of their virion structure, assembly and associated functions remain to be unveiled. In this study, we present a comprehensive mutational and molecular analysis of the temperate Lactococcus lactis-infecting phage TP901-1. Fourteen mutations located within the structural module of TP901-1 were created; twelve mutations were designed to prevent full length translation of putative proteins by non-sense mutations, while two additional mutations caused aberrant protein production. Electron microscopy and Western blot analysis of mutant virion preparations, as well as in vitro assembly of phage mutant combinations, revealed the essential nature of many of the corresponding gene products and provided information on their biological function(s). Based on the information obtained, we propose a functional and assembly model of the TP901-1 Siphoviridae virion.

No MeSH data available.


Related in: MedlinePlus