Molecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP27

ABSTRACT Serine-arginine (SR) protein kinase 1 (SRPK1) catalyzes the phosphorylation of SR proteins, which are a conserved family of splicing factors that contain a domain rich in arginine and serine repeats. SR proteins play important roles in constitutive pre-mRNA splicing and are also important r...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Richard B. Tunnicliffe, William K. Hu, Michele Y. Wu, Colin Levy, A. Paul Mould, Edward A. McKenzie, Rozanne M. Sandri-Goldin, Alexander P. Golovanov
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	phosphorylation splicing herpes simplex virus structure SR protein kinase Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/a3d1cf2d127e48568e7e7049c2934cf3
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a3d1cf2d127e48568e7e7049c2934cf3
record_format	dspace
spelling	oai:doaj.org-article:a3d1cf2d127e48568e7e7049c2934cf32021-11-15T15:59:42ZMolecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP2710.1128/mBio.02551-192150-7511https://doaj.org/article/a3d1cf2d127e48568e7e7049c2934cf32019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02551-19https://doaj.org/toc/2150-7511ABSTRACT Serine-arginine (SR) protein kinase 1 (SRPK1) catalyzes the phosphorylation of SR proteins, which are a conserved family of splicing factors that contain a domain rich in arginine and serine repeats. SR proteins play important roles in constitutive pre-mRNA splicing and are also important regulators of alternative splicing. During herpes simplex virus infection, SRPK1 is inactivated and its cellular distribution is markedly altered by interaction with the viral protein ICP27, resulting in hypophosphorylation of SR proteins. Mutational analysis previously showed that the RGG box motif of ICP27 is required for interaction with SRPK1; however, the mechanism for the inhibition and the exact role of the RGG box was unknown. Here, we used solution nuclear magnetic resonance (NMR) spectroscopy and isothermal titration calorimetry (ITC) to demonstrate that the isolated peptide comprising the RGG box of ICP27 binds to SRPK1 with high affinity, competing with a native substrate, the SR repeat region of SR protein SRSF1. We determined the crystal structure of the complex between SRPK1 and an RGG box peptide, which revealed that the viral peptide binds to the substrate docking groove, mimicking the interactions of SR repeats. Site-directed mutagenesis within the RGG box further confirmed the importance of selected arginine residues for interaction, relocalization, and inhibition of SRPK1 in vivo. Together these data reveal the molecular mechanism of the competitive inhibition of cellular SRPK1 by viral ICP27, which modulates SRPK1 activity. IMPORTANCE Serine arginine (SR) proteins are a family of mRNA regulatory proteins that can modulate spliceosome association with different splice sites and therefore regulate alternative splicing. Phosphorylation within SR proteins is necessary for splice-site recognition, and this is catalyzed by SR protein kinase 1 (SRPK1). The herpes simplex virus (HSV-1) protein ICP27 has been shown previously to interact with and downregulate SRPK1 activity in vivo; however, the molecular mechanism for this interaction and inhibition was unknown. Here, we demonstrate that the isolated peptide fragment of ICP27 containing RGG box binds to SRPK1 with high affinity, and competes with a native cellular substrate. Elucidation of the SRPK1-RGG box crystal structure further showed that a short palindromic RGRRRGR sequence binds in the substrate docking groove of SRPK1, mimicking the binding of SR repeats of substrates. These data reveal how the viral protein ICP27 inactivates SRPK1, promoting hypophosphorylation of proteins regulating splicing.Richard B. TunnicliffeWilliam K. HuMichele Y. WuColin LevyA. Paul MouldEdward A. McKenzieRozanne M. Sandri-GoldinAlexander P. GolovanovAmerican Society for Microbiologyarticlephosphorylationsplicingherpes simplex virusstructurephosphorylationSR protein kinaseMicrobiologyQR1-502ENmBio, Vol 10, Iss 5 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	phosphorylation splicing herpes simplex virus structure phosphorylation SR protein kinase Microbiology QR1-502
spellingShingle	phosphorylation splicing herpes simplex virus structure phosphorylation SR protein kinase Microbiology QR1-502 Richard B. Tunnicliffe William K. Hu Michele Y. Wu Colin Levy A. Paul Mould Edward A. McKenzie Rozanne M. Sandri-Goldin Alexander P. Golovanov Molecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP27
description	ABSTRACT Serine-arginine (SR) protein kinase 1 (SRPK1) catalyzes the phosphorylation of SR proteins, which are a conserved family of splicing factors that contain a domain rich in arginine and serine repeats. SR proteins play important roles in constitutive pre-mRNA splicing and are also important regulators of alternative splicing. During herpes simplex virus infection, SRPK1 is inactivated and its cellular distribution is markedly altered by interaction with the viral protein ICP27, resulting in hypophosphorylation of SR proteins. Mutational analysis previously showed that the RGG box motif of ICP27 is required for interaction with SRPK1; however, the mechanism for the inhibition and the exact role of the RGG box was unknown. Here, we used solution nuclear magnetic resonance (NMR) spectroscopy and isothermal titration calorimetry (ITC) to demonstrate that the isolated peptide comprising the RGG box of ICP27 binds to SRPK1 with high affinity, competing with a native substrate, the SR repeat region of SR protein SRSF1. We determined the crystal structure of the complex between SRPK1 and an RGG box peptide, which revealed that the viral peptide binds to the substrate docking groove, mimicking the interactions of SR repeats. Site-directed mutagenesis within the RGG box further confirmed the importance of selected arginine residues for interaction, relocalization, and inhibition of SRPK1 in vivo. Together these data reveal the molecular mechanism of the competitive inhibition of cellular SRPK1 by viral ICP27, which modulates SRPK1 activity. IMPORTANCE Serine arginine (SR) proteins are a family of mRNA regulatory proteins that can modulate spliceosome association with different splice sites and therefore regulate alternative splicing. Phosphorylation within SR proteins is necessary for splice-site recognition, and this is catalyzed by SR protein kinase 1 (SRPK1). The herpes simplex virus (HSV-1) protein ICP27 has been shown previously to interact with and downregulate SRPK1 activity in vivo; however, the molecular mechanism for this interaction and inhibition was unknown. Here, we demonstrate that the isolated peptide fragment of ICP27 containing RGG box binds to SRPK1 with high affinity, and competes with a native cellular substrate. Elucidation of the SRPK1-RGG box crystal structure further showed that a short palindromic RGRRRGR sequence binds in the substrate docking groove of SRPK1, mimicking the binding of SR repeats of substrates. These data reveal how the viral protein ICP27 inactivates SRPK1, promoting hypophosphorylation of proteins regulating splicing.
format	article
author	Richard B. Tunnicliffe William K. Hu Michele Y. Wu Colin Levy A. Paul Mould Edward A. McKenzie Rozanne M. Sandri-Goldin Alexander P. Golovanov
author_facet	Richard B. Tunnicliffe William K. Hu Michele Y. Wu Colin Levy A. Paul Mould Edward A. McKenzie Rozanne M. Sandri-Goldin Alexander P. Golovanov
author_sort	Richard B. Tunnicliffe
title	Molecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP27
title_short	Molecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP27
title_full	Molecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP27
title_fullStr	Molecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP27
title_full_unstemmed	Molecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP27
title_sort	molecular mechanism of sr protein kinase 1 inhibition by the herpes virus protein icp27
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/a3d1cf2d127e48568e7e7049c2934cf3
work_keys_str_mv	AT richardbtunnicliffe molecularmechanismofsrproteinkinase1inhibitionbytheherpesvirusproteinicp27 AT williamkhu molecularmechanismofsrproteinkinase1inhibitionbytheherpesvirusproteinicp27 AT micheleywu molecularmechanismofsrproteinkinase1inhibitionbytheherpesvirusproteinicp27 AT colinlevy molecularmechanismofsrproteinkinase1inhibitionbytheherpesvirusproteinicp27 AT apaulmould molecularmechanismofsrproteinkinase1inhibitionbytheherpesvirusproteinicp27 AT edwardamckenzie molecularmechanismofsrproteinkinase1inhibitionbytheherpesvirusproteinicp27 AT rozannemsandrigoldin molecularmechanismofsrproteinkinase1inhibitionbytheherpesvirusproteinicp27 AT alexanderpgolovanov molecularmechanismofsrproteinkinase1inhibitionbytheherpesvirusproteinicp27
_version_	1718426987294359552

Molecular Mechanism of SR Protein Kinase 1 Inhibition by the Herpes Virus Protein ICP27

Ejemplares similares