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Artamonova, T.

Publications and source records attributed to Artamonova, T..

2 recordsLinked to original sources

The Dynamics of Synthesis and Localization of Jumbo Phage RNA Polymerases inside Infected Cells

A nucleus-like structure composed of phage-encoded proteins and containing replicating viral DNA is formed in Pseudomonas aeruginosa cells infected by jumbo bacteriophage phiKZ. The PhiKZ genes are transcribed independently from host RNA polymerase (RNAP) by two RNAPs encoded by the phage. The virion RNAP (vRNAP) transcribes early viral genes and must be injected into the cell with phage DNA. The non-virion RNAP (nvRNAP) is composed of early genes products and transcribes late viral genes. In this work, the dynamics of phage RNAPs localization during phage phiKZ infection was studied. We provide direct evidence of PhiKZ vRNAP injection in infected cells and show that it is excluded from the phage nucleus. The nvRNAP is synthesized shortly after the onset of infection and localizes in the nucleus. We propose that spatial separation of two phage RNAPs allows coordinated expression of phage genes belonging to different temporal classes.

molecular biology↗

Structure of the Bacteriophage PhiKZ non-Virion RNA Polymerase

Bacteriophage {Phi}KZ is the founding member of a family of massive bacterial viruses. It is considered to have therapeutic potential as its host, Pseudomonas aeruginosa, is an opportunistic, intrinsically antibiotic resistant, pathogen that kills tens of thousands worldwide each year. {Phi}KZ is an incredibly interesting virus, expressing many systems the host already possesses. On infection, it forms a "nucleus", erecting a barrier around its massive genome to exclude host restriction endonucleases and CRISPR-Cas systems. {Phi}KZ infection is independent of the host transcriptional apparatus. It expresses two different multi-subunit RNA polymerases (RNAPs): the virion RNAP (vRNAP) is injected with the viral DNA during infection to transcribe early genes, including those encoding the non-virion RNAP (nvRNAP), which transcribes all further genes. {Phi}KZ nvRNAP is formed by four polypeptides thought to represent homologues of the eubacterial {beta}/{beta}' subunits, and a fifth with unclear homology, but essential for transcription. We have resolved the structure of {Phi}KZ nvRNAP to 3.3 [A], shedding light on its assembly, homology, and the biological role of the fifth subunit: it is an embedded, integral member of the complex, with structural homology and a biochemical role implying that it has evolved from an ancestral homologue to {sigma}-factor.

biochemistry↗