Human Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP35

The human parainfluenza virus 3 (HPIV3) poses a risk for pneumonia development in young children and immunocompromised patients. To investigate mechanisms of HPIV3 pathogenesis, we characterized the association state and host protein interactions of HPIV3 phosphoprotein (HPIV3 P), an indispensable v...

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Autores principales:	Joaquin Rodriguez Galvan, Brianna Donner, Cat Hoang Veseley, Patrick Reardon, Heather M. Forsythe, Jesse Howe, Gretchen Fujimura, Elisar Barbar
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	HPIV3 EBOV phosphoprotein intrinsically disordered proteins VP35 LC8 Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ed76d39d20b4412ea6254cf810f84d87
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spelling	oai:doaj.org-article:ed76d39d20b4412ea6254cf810f84d872021-11-25T16:52:43ZHuman Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP3510.3390/biom111116032218-273Xhttps://doaj.org/article/ed76d39d20b4412ea6254cf810f84d872021-10-01T00:00:00Zhttps://www.mdpi.com/2218-273X/11/11/1603https://doaj.org/toc/2218-273XThe human parainfluenza virus 3 (HPIV3) poses a risk for pneumonia development in young children and immunocompromised patients. To investigate mechanisms of HPIV3 pathogenesis, we characterized the association state and host protein interactions of HPIV3 phosphoprotein (HPIV3 P), an indispensable viral polymerase cofactor. Sequence analysis and homology modeling predict that HPIV3 P possesses a long, disordered N-terminal tail (P<sub>TAIL</sub>) a coiled-coil multimerization domain (P<sub>MD</sub>), similar to the well-characterized paramyxovirus phosphoproteins from measles and Sendai viruses. Using a recombinantly expressed and purified construct of P<sub>MD</sub> and P<sub>TAIL</sub>, we show that HPIV3 P in solution is primarily an alpha-helical tetramer that is stable up to 60 °C. Pulldown and isothermal titration calorimetry experiments revealed that HPIV3 P binds the host hub protein LC8, and turbidity experiments demonstrated a new role for LC8 in increasing the solubility of HPIV3 P in the presence of crowding agents such as RNA. For comparison, we show that the multimerization domain of the Zaire Ebola virus phosphoprotein VP35 is also a tetramer and binds LC8 but with significantly higher affinity. Comparative analysis of the domain architecture of various virus phosphoproteins in the order Mononegavirales show multiple predicted and verified LC8 binding motifs, suggesting its prevalence and importance in regulating viral phosphoprotein structures. Our work provides evidence for LC8 binding to phosphoproteins with multiple association states, either tetrameric, as in the HPIV3 and Ebola phosphoproteins shown here, or dimeric as in rabies virus phosphoprotein. Taken together the data suggest that the association states of a virus-specific phosphoprotein and the complex formed by binding of the phosphoprotein to host LC8 are important regulators of viral function.Joaquin Rodriguez GalvanBrianna DonnerCat Hoang VeseleyPatrick ReardonHeather M. ForsytheJesse HoweGretchen FujimuraElisar BarbarMDPI AGarticleHPIV3EBOVphosphoproteinintrinsically disordered proteinsVP35LC8MicrobiologyQR1-502ENBiomolecules, Vol 11, Iss 1603, p 1603 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	HPIV3 EBOV phosphoprotein intrinsically disordered proteins VP35 LC8 Microbiology QR1-502
spellingShingle	HPIV3 EBOV phosphoprotein intrinsically disordered proteins VP35 LC8 Microbiology QR1-502 Joaquin Rodriguez Galvan Brianna Donner Cat Hoang Veseley Patrick Reardon Heather M. Forsythe Jesse Howe Gretchen Fujimura Elisar Barbar Human Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP35
description	The human parainfluenza virus 3 (HPIV3) poses a risk for pneumonia development in young children and immunocompromised patients. To investigate mechanisms of HPIV3 pathogenesis, we characterized the association state and host protein interactions of HPIV3 phosphoprotein (HPIV3 P), an indispensable viral polymerase cofactor. Sequence analysis and homology modeling predict that HPIV3 P possesses a long, disordered N-terminal tail (P<sub>TAIL</sub>) a coiled-coil multimerization domain (P<sub>MD</sub>), similar to the well-characterized paramyxovirus phosphoproteins from measles and Sendai viruses. Using a recombinantly expressed and purified construct of P<sub>MD</sub> and P<sub>TAIL</sub>, we show that HPIV3 P in solution is primarily an alpha-helical tetramer that is stable up to 60 °C. Pulldown and isothermal titration calorimetry experiments revealed that HPIV3 P binds the host hub protein LC8, and turbidity experiments demonstrated a new role for LC8 in increasing the solubility of HPIV3 P in the presence of crowding agents such as RNA. For comparison, we show that the multimerization domain of the Zaire Ebola virus phosphoprotein VP35 is also a tetramer and binds LC8 but with significantly higher affinity. Comparative analysis of the domain architecture of various virus phosphoproteins in the order Mononegavirales show multiple predicted and verified LC8 binding motifs, suggesting its prevalence and importance in regulating viral phosphoprotein structures. Our work provides evidence for LC8 binding to phosphoproteins with multiple association states, either tetrameric, as in the HPIV3 and Ebola phosphoproteins shown here, or dimeric as in rabies virus phosphoprotein. Taken together the data suggest that the association states of a virus-specific phosphoprotein and the complex formed by binding of the phosphoprotein to host LC8 are important regulators of viral function.
format	article
author	Joaquin Rodriguez Galvan Brianna Donner Cat Hoang Veseley Patrick Reardon Heather M. Forsythe Jesse Howe Gretchen Fujimura Elisar Barbar
author_facet	Joaquin Rodriguez Galvan Brianna Donner Cat Hoang Veseley Patrick Reardon Heather M. Forsythe Jesse Howe Gretchen Fujimura Elisar Barbar
author_sort	Joaquin Rodriguez Galvan
title	Human Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP35
title_short	Human Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP35
title_full	Human Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP35
title_fullStr	Human Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP35
title_full_unstemmed	Human Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP35
title_sort	human parainfluenza virus 3 phosphoprotein is a tetramer and shares structural and interaction features with ebola phosphoprotein vp35
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/ed76d39d20b4412ea6254cf810f84d87
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Human Parainfluenza Virus 3 Phosphoprotein Is a Tetramer and Shares Structural and Interaction Features with Ebola Phosphoprotein VP35

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