Comprehensive Mutational Analysis of a Herpesvirus Gene in the Viral Genome Context Reveals a Region Essential for Virus Replication

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 2004 Jul 16

PMID 15254174

Citations 54

Authors

Anja Bubeck

Markus Wagner

Zsolt Ruzsics

Mark Lotzerich

Margot Iglesias

Ila R Singh

Ulrich H Koszinowski

Affiliations

Soon will be listed here.

Abstract

Essential viral proteins perform vital functions during morphogenesis via a complex interaction with other viral and cellular gene products. Here, we present a novel approach to comprehensive mutagenesis of essential cytomegalovirus genes and biological analysis in the 230-kbp-genome context. A random Tn7-based mutagenesis procedure at the single-gene level was combined with site-specific recombination via the FLP/FLP recognition target site system for viral genome reconstitution. We show the function of more than 100 mutants from a larger library of M50/p35, a protein involved in capsid egress from the nucleus. This protein recruits other viral proteins and cellular enzymes to the inner nuclear membrane. Our approach enabled us to rapidly discriminate between essential and nonessential regions within the coding sequence. Based on the prediction of the screen, we were able to map a site essential for viral protein-protein interaction at the amino acid level.

Citing Articles

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