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A phage nucleus-associated RNA-binding protein is required for jumbo phage infection.

Nucleic acids research · 2024 · Vol. 52 (8) · pp. 4440-4455

Abstract

Large-genome bacteriophages (jumbo phages) of the proposed family Chimalliviridae assemble a nucleus-like compartment bounded by a protein shell that protects the replicating phage genome from host-encoded restriction enzymes and DNA-targeting CRISPR-Cas nucleases. While the nuclear shell provides broad protection against host nucleases, it necessitates transport of mRNA out of the nucleus-like compartment for translation by host ribosomes, and transport of specific proteins into the nucleus-like compartment to support DNA replication and mRNA transcription. Here, we identify a conserved phage nuclear shell-associated protein that we term Chimallin C (ChmC), which adopts a nucleic acid-binding fold, binds RNA with high affinity in vitro, and binds phage mRNAs in infected cells. ChmC also forms phase-separated condensates with RNA in vitro. Targeted knockdown of ChmC using mRNA-targeting dCas13d results in accumulation of phage-encoded mRNAs in the phage nucleus, reduces phage protein production, and compromises virion assembly. Taken together, our data show that the conserved ChmC protein plays crucial roles in the viral life cycle, potentially by facilitating phage mRNA translocation through the nuclear shell to promote protein production and virion development.

Publication Types

["Journal Article", "Research Support, Non-U.S. Gov't", "Research Support, N.I.H., Extramural"]

Keywords

[]

MeSH Terms

["Bacteriophages", "Cell Nucleus", "CRISPR-Cas Systems", "Genome, Viral", "RNA, Messenger", "RNA, Viral", "RNA-Binding Proteins", "Viral Proteins", "Virus Assembly"]

Funding

R35 GM144121 NIGMS NIH HHS (United States)
R01 GM129245 NIH HHS (United States)
S10 OD021724 NIH HHS (United States)
Howard Hughes Medical Institute (United States)
R01 GM129245 NIGMS NIH HHS (United States)

Linked Datasets (1)

GSE243675 GSE via ncbi_elink
GEO

A phage nucleus-associated RNA-binding protein required for jumbo phage infection

287
16 data files
FileTypeSize
SA_gp61-sfGFP_IN_1_S7_L002_R1_001.fastq.gz RNA-Seq 682.9 MB
SA_gp61-sfGFP_IN_1_S7_L002_R1_001.fastq.gz RNA-Seq 682.9 MB
SA_gp61-sfGFP_IN_2_S5_L002_R1_001.fastq.gz RNA-Seq 640.0 MB
SA_gp61-sfGFP_IN_2_S5_L002_R1_001.fastq.gz RNA-Seq 640.0 MB
SA_gp61-sfGFP_IP_1_S11_L002_R1_001.fastq.gz RIP-Seq 588.1 MB
SA_gp61-sfGFP_IP_1_S11_L002_R1_001.fastq.gz RIP-Seq 588.1 MB
SA_gp61-sfGFP_IP_2_S9_L002_R1_001.fastq.gz RIP-Seq 392.2 MB
SA_gp61-sfGFP_IP_2_S9_L002_R1_001.fastq.gz RIP-Seq 392.2 MB
SA_sfGFP_IN_1_S4_L002_R1_001.fastq.gz RNA-Seq 476.7 MB
SA_sfGFP_IN_1_S4_L002_R1_001.fastq.gz RNA-Seq 476.7 MB
SA_sfGFP_IN_2_S6_L002_R1_001.fastq.gz RNA-Seq 551.8 MB
SA_sfGFP_IN_2_S6_L002_R1_001.fastq.gz RNA-Seq 551.8 MB
SA_sfGFP_IP_1_S8_L002_R1_001.fastq.gz RIP-Seq 272.2 MB
SA_sfGFP_IP_1_S8_L002_R1_001.fastq.gz RIP-Seq 272.2 MB
SA_sfGFP_IP_2_S10_L002_R1_001.fastq.gz RIP-Seq 604.7 MB
SA_sfGFP_IP_2_S10_L002_R1_001.fastq.gz RIP-Seq 604.7 MB

Potentially Related Datasets (2)

These accessions were text-mined from the PMC full text. They may be referenced for comparison, cited from other studies, or otherwise mentioned without being primary data for this paper.

A phage nucleus-associated RNA-binding protein required for jumbo phage infection

PXD045260 PXD PRIDE

Analysis Pipelines (1)

eCLIP geo_data_processing GSE243675