A biochemical portrait of the nidovirus RNA polymerases and helicase

This thesis discusses the purification and activities of the SARS-coronavirus (SARS-CoV) RNA-dependent RNA polymerases (RdRps) nsp12 and nsp(7+8). The first is a large monomeric RdRp, whose stability is greatly influenced by N-terminal additions. In contrast, the latter RdRp is a remarkable hexadecameric complex that is capable of both de novo initiation and primer-extension. In addition these RNA polymerase activities, the thesis also expatiates upon the calibration of magnetic tweezers for force measurements and the dynamics of the equine arteritis virus (EAV) RNA helicase as function of NTP hydrolysis.For financial support, the Netherlands Organization for Scientific Research (NWO) is gratefully acknowledged for Toptalent grant 021.001.037 and Open Access grants 036.001.008, 036.001.250, and 036.001.619 for the publication of chapters 3, 4 and 5.UBL - phd migration 201

A biochemical portrait of the nidovirus RNA polymerases and helicase

This thesis discusses the purification and activities of the SARS-coronavirus (SARS-CoV) RNA-dependent RNA polymerases (RdRps) nsp12 and nsp(7+8). The first is a large monomeric RdRp, whose stability is greatly influenced by N-terminal additions. In contrast, the latter RdRp is a remarkable hexadecameric complex that is capable of both de novo initiation and primer-extension. In addition these RNA polymerase activities, the thesis also expatiates upon the calibration of magnetic tweezers for force measurements and the dynamics of the equine arteritis virus (EAV) RNA helicase as function of NTP hydrolysis