The discovery of small RNA species that are involved in many bacterial regulatory processes (Repolia & Gottesman, 2003; Gottesman et al., 2006; Marles-Wright & Lewis, 2007) supports this possibility. Further studies
on RNA products resulting from the RNase III cleavage of mRNA are needed to address this possibility. This research was supported by grants from the National Research Foundation of Korea (NRF-2009-0065181 and NRF-2010-0029167). K.K. and S.-H.S. equally contributed to this work. Table S1. Analysis of bdm loop mutants. Please note: Wiley-Blackwell is not responsible for the content or functionality BGB324 in vivo of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“A gene product of ORF24′ was identified on the genome of corynephage BFK20 as a putative phage endolysin. The protein of endolysin BFK20 (gp24′) has a modular structure consisting of an N-terminal amidase_2 domain (gp24CD) and a C-terminal cell wall binding domain (gp24BD). The C-terminal domain
is unrelated to any of the known cell wall binding domains of phage endolysins. The whole endolysin gene and the sequences of its N-terminal and C-terminal domains were cloned; proteins were expressed in Escherichia coli and purified to homogeneity. The lytic activities of endolysin and its catalytic domain were demonstrated on corynebacteria selleck inhibitor and bacillus substrates. The binding activity of cell wall binding domain alone and in fusion with green fluorescent protein (gp24BD-GFP) were shown by specific binding assays to the cell surface of BFK20 host Brevibacterium flavum CCM 251 as well as those of other corynebacteria. Phage endolysins Inositol monophosphatase 1 hydrolyze the cell wall of host bacteria from within to release phage progeny at the end of the bacteriophage lytic cycle. Most endolysins
need the help of another phage protein named holin. This small transmembrane protein creates pores in the cytoplasmatic membrane and enables endolysin to pass through the membrane into the periplasma to reach its substrate, a cell wall peptidoglycan. Endolysins of phages that infect Gram-negative bacteria are mostly single-domain globular proteins (Briers et al., 2007). Endolysins isolated from phages targeting Gram-positive bacteria are reported mostly as multidomain structures possessing at least two distinct functional domains, an N-terminal catalytic domain and a C-terminal cell wall binding domain (Loessner, 2005; Fischetti, 2010). The catalytic domain is responsible for the muralytic activities directed against the three different covalent linkages that maintain the integrity of the cell wall. Endolysins have been divided into five classes based on their enzymatic specificity; most of them are amidases and muramidases (Loessner, 2005).