TY - JOUR
T1 - Molecular modelling of the HCMV IL-10 protein isoforms and analysis of their interaction with the human IL-10 receptor
AU - Pantaleão, Simone Queiroz
AU - Camillo, Lívia de Moraes Bomediano
AU - Neves, Tainan Cerqueira
AU - Menezes, Isabela de Godoy
AU - Stangherlin, Lucas Matheus
AU - Batista, Helena Beatriz de Carvalho Ruthner
AU - Poole, Emma
AU - Nevels, Michael
AU - Philot, Eric Alisson
AU - Scott, Ana Ligia
AU - Carlan da Silva, Maria Cristina
N1 - Funding: Funder 01: Grant number: 2020/08527-1 Fundação de Amparo a Pesquisa do Estado de São Paulo, Brazil (FAPESP) - https://fapesp.br/ Author: Maria Cristina Carlan da Silva - Maria Cristina Carlan Silva Funder 02: Grant number: 2020/07767-9 Fundação de Amparo a Pesquisa do Estado de São Paulo, Brazil (FAPESP) - https://fapesp.br/ Author: Tainan Cerqueira Neves - Neves, Tainan C. Funder 03: Grant number: 164052/2020-8 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) - https://www.gov.br/cnpq/pt-br Author: Simone Queiroz Pantaleão - Pantaleão, Simone Queiroz.
PY - 2022/11/28
Y1 - 2022/11/28
N2 - The human cytomegalovirus (HCMV) UL111A gene encodes several homologs of the cellular interleukin 10 (cIL-10). Alternative splicing in the UL111A region produces two relatively well-characterized transcripts designated cmvIL-10 (isoform A) and LAcmvIL-10 (isoform B). The cmvIL-10 protein is the best characterized, both structurally and functionally, and has many immunosuppressive activities similar to cIL-10, while LAcmvIL-10 has more restricted biological activities. Alternative splicing also results in five less studied UL111A transcripts encoding additional proteins homologous to cIL-10 (isoforms C to G). These transcripts were identified during productive HCMV infection of MRC-5 cells with the high passage laboratory adapted AD169 strain, and the structure and properties of the corresponding proteins are largely unknown. Moreover, it is unclear whether these protein isoforms are able to bind the cellular IL-10 receptor and induce signalling. In the present study, we investigated the expression spectrum of UL111A transcripts in fully permissive MRC-5 cells and semi permissive U251 cells infected with the low passage HCMV strain TB40E. We identified a new spliced transcript (H) expressed during productive infection. Using computational methods, we carried out molecular modelling studies on the three-dimensional structures of the HCMV IL-10 proteins encoded by the transcripts detected in our work (cmvIL-10 (A), LAcmvIL-10 (B), E, F and H) and on their interaction with the human IL-10 receptor (IL-10R1). The modelling predicts clear differences between the isoform structures. Furthermore, the in silico simulations (molecular dynamics simulation and normal-mode analyses) allowed us to evaluate regions that contain potential receptor binding sites in each isoform. The analyses demonstrate that the complexes between the isoforms and IL-10R1 present different types of molecular interactions and consequently different affinities and stabilities. The knowledge about structure and expression of specific viral IL-10 isoforms has implications for understanding of their properties and role in HCMV immune evasion and pathogenesis.
AB - The human cytomegalovirus (HCMV) UL111A gene encodes several homologs of the cellular interleukin 10 (cIL-10). Alternative splicing in the UL111A region produces two relatively well-characterized transcripts designated cmvIL-10 (isoform A) and LAcmvIL-10 (isoform B). The cmvIL-10 protein is the best characterized, both structurally and functionally, and has many immunosuppressive activities similar to cIL-10, while LAcmvIL-10 has more restricted biological activities. Alternative splicing also results in five less studied UL111A transcripts encoding additional proteins homologous to cIL-10 (isoforms C to G). These transcripts were identified during productive HCMV infection of MRC-5 cells with the high passage laboratory adapted AD169 strain, and the structure and properties of the corresponding proteins are largely unknown. Moreover, it is unclear whether these protein isoforms are able to bind the cellular IL-10 receptor and induce signalling. In the present study, we investigated the expression spectrum of UL111A transcripts in fully permissive MRC-5 cells and semi permissive U251 cells infected with the low passage HCMV strain TB40E. We identified a new spliced transcript (H) expressed during productive infection. Using computational methods, we carried out molecular modelling studies on the three-dimensional structures of the HCMV IL-10 proteins encoded by the transcripts detected in our work (cmvIL-10 (A), LAcmvIL-10 (B), E, F and H) and on their interaction with the human IL-10 receptor (IL-10R1). The modelling predicts clear differences between the isoform structures. Furthermore, the in silico simulations (molecular dynamics simulation and normal-mode analyses) allowed us to evaluate regions that contain potential receptor binding sites in each isoform. The analyses demonstrate that the complexes between the isoforms and IL-10R1 present different types of molecular interactions and consequently different affinities and stabilities. The knowledge about structure and expression of specific viral IL-10 isoforms has implications for understanding of their properties and role in HCMV immune evasion and pathogenesis.
U2 - 10.1371/journal.pone.0277953
DO - 10.1371/journal.pone.0277953
M3 - Article
SN - 1932-6203
VL - 17
JO - PLoS ONE
JF - PLoS ONE
IS - 11
M1 - e0277953
ER -