2MML

T47 phosphorylation of the Mengovirus Leader Protein: NMR Studies of the Phosphorylation of the Mengovirus Leader Protein Reveal Stabilization of Intermolecular Domain Interactions

Summary for 2MML

• Entry DOI: 10.2210/pdb2mml/pdb
• Related: 2MMG 2MMH 2MMI 2MMK
• NMR Information: BMRB: 19858
• Descriptor: Leader protein, ZINC ION (2 entities in total)
• Functional Keywords: animal viruses, positive-strand rna viruses, cardioviruses, leader, protein phosphorylation, casein kinase 2, viral protein
• Biological source: Mengo virus
• Cellular location: Protein 2A: Host nucleus . Protein VP2: Virion. Protein VP3: Virion. Protein VP1: Virion. Protein 2B: Host cytoplasmic vesicle membrane ; Peripheral membrane protein ; Cytoplasmic side . Protein 2C: Host cytoplasmic vesicle membrane ; Peripheral membrane protein ; Cytoplasmic side . Protein 3A: Host cytoplasmic vesicle membrane ; Peripheral membrane protein ; Cytoplasmic side . Protein 3B: Virion . Protease 3C: Host cytoplasm . RNA-directed RNA polymerase 3D-POL: Host cytoplasmic vesicle membrane ; Peripheral membrane protein ; Cytoplasmic side : P12296
• Total number of polymer chains: 1
• Total formula weight: 8407.43

Authors

Bacot-Davis, V.R.,Porter, F.W.,Palmenberg, A.C. (deposition date: 2014-03-15, release date: 2014-10-15, Last modification date: 2024-10-30)

Primary citation

Bacot-Davis, V.R.,Ciomperlik, J.J.,Basta, H.A.,Cornilescu, C.C.,Palmenberg, A.C.
Solution structures of Mengovirus Leader protein, its phosphorylated derivatives, and in complex with nuclear transport regulatory protein, RanGTPase.
Proc.Natl.Acad.Sci.USA, 111:15792-15797, 2014

PubMed Abstract: Cardiovirus Leader (L) proteins induce potent antihost inhibition of active cellular nucleocytoplasmic trafficking by triggering aberrant hyperphosphorylation of nuclear pore proteins (Nup). To achieve this, L binds protein RanGTPase (Ran), a key trafficking regulator, and diverts it into tertiary or quaternary complexes with required kinases. The activity of L is regulated by two phosphorylation events not required for Ran binding. Matched NMR studies on the unphosphorylated, singly, and doubly phosphorylated variants of Mengovirus L (L(M)) show both modifications act together to partially stabilize a short internal α-helix comprising L(M) residues 43-46. This motif implies that ionic and Van der Waals forces contributed by phosphorylation help organize downstream residues 48-67 into a new interface. The full structure of L(M) as bound to Ran (unlabeled) and Ran (216 aa) as bound by L(M) (unlabeled) places L(M) into the BP1 binding site of Ran, wrapped by the conformational flexible COOH tail. The arrangement explains the tight KD for this complex and places the LM zinc finger and phosphorylation interface as surface exposed and available for subsequent reactions. The core structure of Ran, outside the COOH tail, is not altered by L(M) binding and remains accessible for canonical RanGTP partner interactions. Pull-down assays identify at least one putative Ran:L(M) partner as an exportin, Crm1, or CAS. A model of Ran:L(M):Crm1, based on the new structures suggests LM phosphorylation status may mediate Ran's selection of exportin(s) and cargo(s), perverting these native trafficking elements into the lethal antihost Nup phosphorylation pathways.

PubMed: 25331866
DOI: 10.1073/pnas.1411098111

Experimental method

SOLUTION NMR