Zinc-binding of the cysteine-rich domain encoded in the open reading frame 1B of the RNA polymerase gene of coronavirus

D. Yoo, M. D. Parker, G. J. Cox, L. A. Babiuk

Research output: Contribution to journalArticlepeer-review

Abstract

We cloned and sequenced the second open reading frame of the RNA polymerase gene, ORF 1b, of bovine coronavirus. In the region representing nucleotide positions 49195677 upstream from the initiation codon of the 32K non-structural protein gene, we identified two putative functional domains. One of these domains contained four leucine residues repeated exactly in every seventh position, and the other domain represented a cluster of cysteine and histidine residues. The DNA sequence representing these domains was cloned and expressed in Escherichia coli as fusion proteins with glutathione S-transferase from Schistosoma japonicum. A high level expression of the cysteine-rich domain was achieved as a fusion protein when the bacterial culture was induced with IPTG. In a solid phase zinc binding assay using the recombinant fusion protein, we found that the protein containing the cysteine-rich domain was able to bind to radioactive zinc in vitro, demonstrating that the polypeptide encoded by the ORF1b of coronavirus is a zinc-binding protein.

Original languageEnglish (US)
Pages (from-to)437-442
Number of pages6
JournalAdvances in experimental medicine and biology
Volume380
DOIs
StatePublished - 1995
Externally publishedYes

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology

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