Unknown Phage 11

Unknown Phage 11Leah Liston

Cluster IdentificationUnknown 11 is identified with Cluster QOther Phage in Cluster QLong time singleton: GilesEvanesceHH92OBUPride

Plaque ObservationsSmall and roundSlightly turbid

Cluster QLightly turbid plaquesSmall

Phamerator - PhamsPham 1278 is unique to Cluster QEvanesce Gene 11 (441bp)Giles Gene 10 (441bp)HH92 Gene 10 (441bp)OBUPride Gene 10 (441bp)The amino acid sequence: (K)AAGAIADASTDEAAAAK(A) At beginning of the genome located after the terminases, protease, portal, and capsid genes

Pham 1278In most phages, between the capsid protein and the tail assembly chaperones proteins there is usually major tail subunit proteinsNo gene in Giles can be predictedPham 1278 protein associated with the unpredicted major tail subunit geneUnique gene because of thisUnknown 11 from Mass Spec had 97% alignmentBiggest Cluster Q indicator

Spectrum MillCompared against phage database top scoresEvanesce gene 11 Pham 1278 Cluster Q specificEvanesce gene 10 Pham 1326 Cluster Q specificEvanesce gene 25 Pham 3145 Cluster Q specificEvanesce gene 16 Pham 1283 Cluster Q specificEvanesce gene 40 Pham 1292 Cluster Q specificBxz2 gene 23 Pham 1842 Not in Cluster QEvanesce gene 9 Pham 1325 Cluster Q specificEvanesce gene 7 Pham 1311 Cluster P & Q onlyDUF935 Unknown function but consists of several bacterial protein including Bacteriophage Mu gp29Bxz2 gene 17 Pham 1841 Not in Cluster Q

6Spectrum MillCompared against Mycobacterium DatabaseEvanesce gene 11 Pham 1278 Cluster Q specificEvanesce gene 10 - Pham 1326 Cluster Q specificElongation factor Tu M. trivaleRNA polymerase beta M. chitaeEndopeptidase Clp 1 MycleGlyceraldehyde-2-phosphate dehydrogenase M. tuberculosis H37RvEvanesce gene 25 Pham 3145 Cluster Q specificATP-dependent Clp protease M. tuberculosis CDC1551Alcohol dehydrogenase M. smegmatis

Spectrum Mill ResultsPhage DatabaseHigh alignments with Evanesce genesHigh alignment with Pham 1278 Cluster Q specificHigh alignment with Cluster Q specific genesMycobacterium DatabaseAlignment with elongation factorCritical function: in translation energy source for binding of tRNA to the A site in ribosomes Endopeptidase Clp 1Major role in degradation of misfolded proteins

Pipeline Comparison with Positive Control Czyszczon1Both ContainElongation Factor Tu Chaperonin GroELGlyceraldehyde-3-phosphate dehydrogenaseFatty acid synthetase I Glutamine synthetase GLNA1Evanesce Gene 22 Pham 3143Only in Cluster Q on Phamerator possible mutationsOnly Unknown 11Glycerol kinaseRNA polymerase betaEvanesce Genes: 17, 43, 11, 10, 21

Chromatography Graphs

Unknown 11Positive Control Czyszcon1

Future DirectionPhameratorLook at cluster Q genes that are shared in other phagesGene Function?

ReferencesHatfull, G. The Secret Lives of Mycobacteriophages. (2012). Advances in Virus ResearchHatfull, G. Comparative genomic analysis of sixty mycobacteriophage genomes: genome clustering, gene acquisition and gene size. (2010). J Mol Biol, 397(1), 119-143Spectrum MillPhameratorNCBI Blastp alignments