TY - JOUR AU - Das, Uddipan AU - Pogenberg, Vivian AU - Tiruttani Subhramanyam, Udaya Kumar AU - Matthias, Wilmanns AU - Gourinath, Samudrala AU - Srinivana, Alagiri TI - Crystal structure of the VapBC-15 complex from Mycobacterium tuberculosis reveals a two-metal ion dependent PIN-domain ribonuclease and a variable mode of toxin-antitoxin assembly JO - Journal of structural biology VL - 188 IS - 3 SN - 1047-8477 CY - San Diego, Calif. PB - Elsevier M1 - FZJ-2014-05967 SP - 249–258 PY - 2014 N1 - Article in press AB - Although PIN (PilT N-terminal)-domain proteins are known to have ribonuclease activity, their specific mechanism of action remains unknown. VapCs form a family of ribonucleases that possess a PIN-domain assembly and are known as toxins. The activities of VapCs are impaired by VapB antitoxins. Here we present the crystal structure of the VapBC-15 toxin–antitoxin complex from Mycobacterium tuberculosis determined to 2.1 Å resolution. The VapB-15 and VapC-15 components assemble into one heterotetramer (VapB2C2) and two heterotrimers (VapBC2) in each asymmetric unit of the crystal. The active site of VapC-15 toxin consists of a cluster of acidic amino acid residues and two divalent metal ions, forming a well organised ribonuclease active site. The distribution of the catalytic-site residues of the VapC-15 toxin is similar to that of T4 RNase H and of Methanococcus jannaschii FEN-1, providing strong evidence that these three proteins share a similar mechanism of activity. The presence of both VapB2C2 and VapBC2 emphasizes the fact that the same antitoxin can bind the toxin in 1:1 and 1:2 ratios. The crystal structure determination of the VapBC-15 complex reveals for the first time a PIN-domain ribonuclease protein that shows two metal ions at the active site and a variable mode of toxin–antitoxin assembly. The structure further shows that VapB-15 antitoxin binds to the same groove meant for the binding of putative substrate (RNA), resulting in the inhibition of VapC-15’s toxicity. LB - PUB:(DE-HGF)16 UR - <Go to ISI:>//WOS:000346229500007 DO - DOI:10.1016/j.jsb.2014.10.002 UR - https://juser.fz-juelich.de/record/172498 ER -