title: The cytotoxic early protein 77 of mycobacteriophage L5 interacts with MSMEG_3532, an L-serine dehydratase of Mycobacterium smegmatis.
abstract: Mycobacteriophage L5 is a temperate phage infecting a broad range of mycobacterial species. Upon induction of lytic growth, L5 rapidly switches off host protein synthesis. We have recently identified the mycobacteriophage L5 early protein gp77 as a host shut-off protein that acts growth inhibitory in the mycobacterial host when expressed through the corresponding phage promoter. Here we present data showing that this purified phage protein of unknown function specifically binds to protein MSMEG_3532 when incubated with cell lysates of Mycobacterium smegmatis. This interaction was confirmed by pull-down assays using purified MSMEG_3532 as bait which co-purified with gp77. The amino acid sequence of MSMEG_3532 is nearly identical to that of threonine dehydratases, serine dehydratases and an L-threo-3-hydroxyaspartate dehydratase. An enzymatic assay identified this host protein as a pyridoxal-5'-phosphate-dependent L-serine dehydratase (SdhA) which converts L-serine to pyruvate. This is the first biochemical characterization of a SdhA derived from mycobacteria. Though the addition of purified gp77 to the established in vitro assay had no influence on SdhA activity at a saturating L-serine concentration, the specific interaction of phage protein and dehydratase in vivo may well have a role in altering the amino acid pool or the products of amino acid metabolism in favour of phage maturation.
authors.affiliation: 1st Department of Internal Medicine, University of Cologne, Cologne, Germany. jan.rybniker@uk-koeln.de
authors.forename: Jan
authors.initials: J
authors.lastname: Rybniker
authors.forename: Karin
authors.initials: K
authors.lastname: Krumbach
authors.forename: Edeltraud
authors.initials: E
authors.lastname: van Gumpel
authors.forename: Georg
authors.initials: G
authors.lastname: Plum
authors.forename: Lothar
authors.initials: L
authors.lastname: Eggeling
authors.forename: Pia
authors.initials: P
authors.lastname: Hartmann
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D003603:Cytotoxins / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005420:Flavoproteins / Q000378:metabolism; D012695:L-Serine Dehydratase / Q000378:metabolism*; D009160:Mycobacteriophages / Q000378:metabolism*; D020102:Mycobacterium smegmatis / Q000201:enzymology* / Q000821:virology; D011485:Protein Binding; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D003603:Cytotoxins; D005420:Flavoproteins; C084271:SdhA protein, Bacteria; D014764:Viral Proteins; D012695:L-Serine Dehydratase