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An essential and highly selective protein import pathway encoded by nucleus-forming phage

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [1];  [1];  [1];  [1];  [3];  [4];  [5];  [6];  [1];  [7];  [8];  [1]
  1. School of Biological Sciences, Division of Molecular Biology, University of California San Diego, La Jolla, CA 92093
  2. School of Biological Sciences, Division of Molecular Biology, University of California San Diego, La Jolla, CA 92093, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand 10330
  3. Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand 10330
  4. California Institute for Quantitative Biosciences, University of California, Berkeley, CA 94720, Innovative Genomics Institute, University of California, Berkeley, CA 94720, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720
  5. Innovative Genomics Institute, University of California, Berkeley, CA 94720, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720
  6. California Institute for Quantitative Biosciences, University of California, Berkeley, CA 94720, Innovative Genomics Institute, University of California, Berkeley, CA 94720, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, Department of Chemistry, University of California, Berkeley, CA 94720, HHMI, University of California, Berkeley, CA 94720
  7. School of Biological Sciences, Division of Molecular Biology, University of California San Diego, La Jolla, CA 92093, HHMI, University of California San Diego, La Jolla, CA 92093
  8. School of Biological Sciences, Division of Molecular Biology, University of California San Diego, La Jolla, CA 92093, Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA 92093

Targeting proteins to specific subcellular destinations is essential in prokaryotes, eukaryotes, and the viruses that infect them. Chimalliviridae phages encapsulate their genomes in a nucleus-like replication compartment composed of the protein chimallin (ChmA) that excludes ribosomes and decouples transcription from translation. These phages selectively partition proteins between the phage nucleus and the bacterial cytoplasm. Currently, the genes and signals that govern selective protein import into the phage nucleus are unknown. Here, we identify two components of this protein import pathway: a species-specific surface-exposed region of a phage intranuclear protein required for nuclear entry and a conserved protein, PicA (Protein importer of chimalliviruses A), that facilitates cargo protein trafficking across the phage nuclear shell. We also identify a defective cargo protein that is targeted to PicA on the nuclear periphery but fails to enter the nucleus, providing insight into the mechanism of nuclear protein trafficking. Using CRISPRi-ART protein expression knockdown of PicA, we show that PicA is essential early in the chimallivirus replication cycle. Together, our results allow us to propose a multistep model for the Protein Import Chimallivirus pathway, where proteins are targeted to PicA by amino acids on their surface and then licensed by PicA for nuclear entry. The divergence in the selectivity of this pathway between closely related chimalliviruses implicates its role as a key player in the evolutionary arms race between competing phages and their hosts.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
National Institutes of Health (NIH); USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
2342165
Alternate ID(s):
OSTI ID: 2407020
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 19 Vol. 121; ISSN 0027-8424
Publisher:
Proceedings of the National Academy of SciencesCopyright Statement
Country of Publication:
United States
Language:
English

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