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TitleStructural Insights into the Roles of PARP4 and NAD in the Human Vault Cage.
Journal, issue, pagesbioRxiv, Year 2024
Publish dateJun 27, 2024
PubMed AbstractVault is a massive ribonucleoprotein complex found across Eukaryota. The major vault protein (MVP) oligomerizes into an ovular cage, which contains several minor vault components (MVCs) and is ...Vault is a massive ribonucleoprotein complex found across Eukaryota. The major vault protein (MVP) oligomerizes into an ovular cage, which contains several minor vault components (MVCs) and is thought to transport transiently bound "cargo" molecules. Vertebrate vaults house a poly (ADP-ribose) polymerase (known as PARP4 in humans), which is the only MVC with known enzymatic activity. Despite being discovered decades ago, the molecular basis for PARP4's interaction with MVP remains unclear. In this study, we determined the structure of the human vault cage in complex with PARP4 and its enzymatic substrate NAD . The structures reveal atomic-level details of the protein-binding interface, as well as unexpected NAD -binding pockets within the interior of the vault cage. In addition, proteomics data show that human vaults purified from wild-type and PARP4-depleted cells interact with distinct subsets of proteins. Our results thereby support a model in which PARP4's specific incorporation into the vault cage helps to regulate vault's selection of cargo and its subcellular localization. Further, PARP4's proximity to MVP's NAD -binding sites could support its enzymatic function within the vault.
External linksbioRxiv / PubMed:38979142 / PubMed Central
MethodsEM (single particle)
Resolution2.68 - 3.4 Å
Structure data

44953

9bw5

EMDB-44953, PDB-9bw5:
Human Vault Cage
Method: EM (single particle) / Resolution: 3.3 Å

44954

9mxh

EMDB-44954, PDB-9mxh:
Human Vault Cage in complex with NAD+
Method: EM (single particle) / Resolution: 3.1 Å

44955

9bw6

EMDB-44955, PDB-9bw6:
Human Vault Cage in complex with PARP4
Method: EM (single particle) / Resolution: 2.9 Å

44957

9bw7

EMDB-44957, PDB-9bw7:
Human Vault Cage in complex with PARP4 and NAD+
Method: EM (single particle) / Resolution: 2.9 Å

44959

9mxv

EMDB-44959: Human Vault Cage in the presence of ADP
PDB-9mxv: Human Vault Cage in complex with ADP
Method: EM (single particle) / Resolution: 2.7 Å

EMDB-44960: Human Vault Cage in complex with the MINT domain of PARP4
Method: EM (single particle) / Resolution: 3.4 Å

Chemicals

ChemComp-NAD:
NICOTINAMIDE-ADENINE-DINUCLEOTIDE / NAD*YM

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

Source
  • homo sapiens (human)
KeywordsPROTEIN TRANSPORT / Vault / Vault Cage / MVP / Major Vault Protein / SPFH / PARP4 / Poly(ADP-ribose)Polymerase 4 / Poly(ADP-ribose)Polymerase / MINT / Ribonucleoprotein / Megadalton complex / TEP1 / vault RNA / Poly(ADP-ribose) Polymerase 4 / NAD+ / NAD / Poly(ADP-ribose) Polymerase / ADP / Adenosine diphosphate

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