7uxe: Difference between revisions
New page: '''Unreleased structure''' The entry 7uxe is ON HOLD Authors: Lokareddy, R.K., Hou, C.-F.D., Doll, S.G., Li, F., Gillilan, R., Forti, F., Briani, F., Cingolani, G. Description: Pseudom... |
No edit summary |
||
| (3 intermediate revisions by the same user not shown) | |||
| Line 1: | Line 1: | ||
==Pseudomonas phage E217 small terminase (TerS)== | |||
<StructureSection load='7uxe' size='340' side='right'caption='[[7uxe]], [[Resolution|resolution]] 3.38Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[7uxe]] is a 10 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_phage_vB_PaeM_E217 Pseudomonas phage vB_PaeM_E217]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7UXE OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7UXE FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 3.38Å</td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=7uxe FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7uxe OCA], [https://pdbe.org/7uxe PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7uxe RCSB], [https://www.ebi.ac.uk/pdbsum/7uxe PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7uxe ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/A0A2K8I4H6_9CAUD A0A2K8I4H6_9CAUD] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Pseudomonas phages are increasingly important biomedicines for phage therapy, but little is known about how these viruses package DNA. This paper explores the terminase subunits from the Myoviridae E217, a Pseudomonas-phage used in an experimental cocktail to eradicate P. aeruginosa in vitro and in animal models. We identified the large (TerL) and small (TerS) terminase subunits in two genes approximately 58 kbs away from each other in the E217 genome. TerL presents a classical two-domain architecture, consisting of an N-terminal ATPase and C-terminal nuclease domain arranged into a bean-shaped tertiary structure. A 2.05 A crystal structure of the C-terminal domain revealed an RNase H-like fold with two magnesium ions in the nuclease active site. Mutations in TerL residues involved in magnesium coordination had a dominant-negative effect on phage growth. However, the two ions identified in the active site were too far from each other to promote two-metal-ion catalysis, suggesting a conformational change is required for nuclease activity. We also determined a 3.38 A cryo-EM reconstruction of E217 TerS that revealed a ring-like decamer, departing from the most common nonameric quaternary structure observed thus far. E217 TerS contains both N-terminal helix-turn-helix motifs enriched in basic residues and a central channel lined with basic residues large enough to accommodate double-stranded DNA. Overexpression of TerS caused a more than a 4-fold reduction of E217 burst size, suggesting a catalytic amount of the protein is required for packaging. Together, these data expand the molecular repertoire of viral terminase subunits to Pseudomonas-phages used for phage therapy. | |||
Terminase Subunits from the Pseudomonas-Phage E217.,Lokareddy RK, Hou CD, Doll SG, Li F, Gillilan RE, Forti F, Horner DS, Briani F, Cingolani G J Mol Biol. 2022 Oct 30;434(20):167799. doi: 10.1016/j.jmb.2022.167799. Epub 2022 , Aug 22. PMID:36007626<ref>PMID:36007626</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
[[Category: | <div class="pdbe-citations 7uxe" style="background-color:#fffaf0;"></div> | ||
[[Category: | |||
[[Category: | ==See Also== | ||
[[Category: | *[[Terminase 3D Structures|Terminase 3D Structures]] | ||
[[Category: Gillilan | == References == | ||
[[Category: | <references/> | ||
[[Category: | __TOC__ | ||
[[Category: | </StructureSection> | ||
[[Category: Large Structures]] | |||
[[Category: Pseudomonas phage vB_PaeM_E217]] | |||
[[Category: Briani F]] | |||
[[Category: Cingolani G]] | |||
[[Category: Doll SG]] | |||
[[Category: Forti F]] | |||
[[Category: Gillilan R]] | |||
[[Category: Hou C-FD]] | |||
[[Category: Li F]] | |||
[[Category: Lokareddy RK]] | |||
Latest revision as of 08:15, 12 June 2024
Pseudomonas phage E217 small terminase (TerS)Pseudomonas phage E217 small terminase (TerS)
Structural highlights
FunctionPublication Abstract from PubMedPseudomonas phages are increasingly important biomedicines for phage therapy, but little is known about how these viruses package DNA. This paper explores the terminase subunits from the Myoviridae E217, a Pseudomonas-phage used in an experimental cocktail to eradicate P. aeruginosa in vitro and in animal models. We identified the large (TerL) and small (TerS) terminase subunits in two genes approximately 58 kbs away from each other in the E217 genome. TerL presents a classical two-domain architecture, consisting of an N-terminal ATPase and C-terminal nuclease domain arranged into a bean-shaped tertiary structure. A 2.05 A crystal structure of the C-terminal domain revealed an RNase H-like fold with two magnesium ions in the nuclease active site. Mutations in TerL residues involved in magnesium coordination had a dominant-negative effect on phage growth. However, the two ions identified in the active site were too far from each other to promote two-metal-ion catalysis, suggesting a conformational change is required for nuclease activity. We also determined a 3.38 A cryo-EM reconstruction of E217 TerS that revealed a ring-like decamer, departing from the most common nonameric quaternary structure observed thus far. E217 TerS contains both N-terminal helix-turn-helix motifs enriched in basic residues and a central channel lined with basic residues large enough to accommodate double-stranded DNA. Overexpression of TerS caused a more than a 4-fold reduction of E217 burst size, suggesting a catalytic amount of the protein is required for packaging. Together, these data expand the molecular repertoire of viral terminase subunits to Pseudomonas-phages used for phage therapy. Terminase Subunits from the Pseudomonas-Phage E217.,Lokareddy RK, Hou CD, Doll SG, Li F, Gillilan RE, Forti F, Horner DS, Briani F, Cingolani G J Mol Biol. 2022 Oct 30;434(20):167799. doi: 10.1016/j.jmb.2022.167799. Epub 2022 , Aug 22. PMID:36007626[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
|
| ||||||||||||||||