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Gene silencing pathway RNA-dependent RNA polymerase of Neurospora crassa: yeast expression, crystallization of selenomethionated QDE-1 protein

Lookup NU author(s): Dr Paula Salgado

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Abstract

The RNA-dependent RNA polymerase, QDE-1, is a component of the RNA silencing pathway in Neurospora crassa. The enzymatically active carboxy-terminal fragment QDE-1 ΔN has been expressed in Saccharomyces cerevisiae in the presence and absence of selenomethionine (SeMet). The level of SeMet incorporation was estimated by mass spectrometry to be ∼98%. Both native and SeMet proteins were crystallized in space group P21 with unit cell parameters a = 101.2, b = 122.5, c = 114.4 Å, β = 108.9°, and 2 molecules per asymmetric unit. The native and SeMet crystals diffract to 2.3 and 3.2 Å, respectively, the latter are suitable for MAD structure determination.


Publication metadata

Author(s): Laurila MR, Salgado PS, Makeyev EV, Nettelship J, Stuart DI, Grimes JM, Bamford DH

Publication type: Article

Publication status: Published

Journal: Journal of Structural Biology

Year: 2005

Volume: 149

Issue: 1

Pages: 111-115

ISSN (print): 1047-8477

ISSN (electronic): 1095-8657

Publisher: Academic Press

URL: http://dx.doi.org/10.1016/j.jsb.2004.10.001

DOI: 10.1016/j.jsb.2004.10.001

PubMed id: 15629662


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