Multivalent dynamic colocalization of avian influenza polymerase and nucleoprotein by intrinsically disordered ANP32A reveals the molecular basis of human adaptation

Camacho-Zarco, A. R.; Yu, L.; Krischuns, T.; Dedeoglu, S.; Maurin, D.; Bouvignies, G.; Crepin, T.; Ruigrok, R. W. H.; Cusack, S.; Naffakh, N.; Blackledge, M.

doi:10.1101/2023.07.08.548184

bioRxiv · 10.1101/2023.07.08.548184

Multivalent dynamic colocalization of avian influenza polymerase and nucleoprotein by intrinsically disordered ANP32A reveals the molecular basis of human adaptation

Camacho-Zarco, A. R.·Yu, L.·Krischuns, T.·Dedeoglu, S.·Maurin, D.·Bouvignies, G.·Crepin, T.·Ruigrok, R. W. H.·Cusack, S.·Naffakh, N.·Blackledge, M.

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Abstract

Adaptation of avian influenza RNA polymerase (FluPol) to human cells requires mutations on the 627-NLS domains of the PB2 subunit. The E627K adaptive mutation compensates a 33-amino-acid deletion in the acidic intrinsically disordered domain of the host transcription regulator ANP32A, a deletion that restricts FluPol activity in mammalian cells. The function of ANP32A in the replication transcription complex and in particular its role in host restriction remain poorly understood. Here we characterise ternary complexes formed between ANP32A, FluPol and the viral nucleoprotein, NP, supporting the putative role of ANP32A in shuttling NP to the replicase complex. We demonstrate that while FluPol and NP can simultaneously bind distinct linear motifs on avian ANP32A, the deletion in the shorter human ANP32A blocks this mode of colocalization. NMR reveals that NP and human-adapted Pol, containing the E627K mutation, simultaneously bind the identical extended linear motif on human ANP32A, in an electrostatically driven, highly dynamic and multivalent ternary complex. This study reveals a probable molecular mechanism underlying host adaptation, whereby E627K, which enhances the basic surface of the 627 domain, is selected to confer the necessary multivalent properties to allow ANP32A to colocalize NP and FluPol in human cells.

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Camacho-Zarco, A. R., Yu, L., Krischuns, T., Dedeoglu, S., Maurin, D., Bouvignies, G., Crepin, T., Ruigrok, R. W. H., Cusack, S., Naffakh, N., Blackledge, M.. 2023-07-08. Multivalent dynamic colocalization of avian influenza polymerase and nucleoprotein by intrinsically disordered ANP32A reveals the molecular basis of human adaptation. https://doi.org/10.1101/2023.07.08.548184

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