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TIM1 (HAVCR1) Is Not Essential for Cellular Entry of Either Quasi-enveloped or Naked Hepatitis A Virions

Receptor molecules play key roles in the cellular entry of picornaviruses, and TIM1 (HAVCR1) is widely accepted to be the receptor for hepatitis A virus (HAV), an unusual, hepatotropic human picornavirus. However, its identification as the hepatovirus receptor predated the discovery that hepatovirus...

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Autores principales: Das, Anshuman, Hirai-Yuki, Asuka, González-López, Olga, Rhein, Bethany, Moller-Tank, Sven, Brouillette, Rachel, Hensley, Lucinda, Misumi, Ichiro, Lovell, William, Cullen, John M., Whitmire, Jason K., Maury, Wendy, Lemon, Stanley M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5587907/
https://www.ncbi.nlm.nih.gov/pubmed/28874468
http://dx.doi.org/10.1128/mBio.00969-17
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author Das, Anshuman
Hirai-Yuki, Asuka
González-López, Olga
Rhein, Bethany
Moller-Tank, Sven
Brouillette, Rachel
Hensley, Lucinda
Misumi, Ichiro
Lovell, William
Cullen, John M.
Whitmire, Jason K.
Maury, Wendy
Lemon, Stanley M.
author_facet Das, Anshuman
Hirai-Yuki, Asuka
González-López, Olga
Rhein, Bethany
Moller-Tank, Sven
Brouillette, Rachel
Hensley, Lucinda
Misumi, Ichiro
Lovell, William
Cullen, John M.
Whitmire, Jason K.
Maury, Wendy
Lemon, Stanley M.
author_sort Das, Anshuman
collection PubMed
description Receptor molecules play key roles in the cellular entry of picornaviruses, and TIM1 (HAVCR1) is widely accepted to be the receptor for hepatitis A virus (HAV), an unusual, hepatotropic human picornavirus. However, its identification as the hepatovirus receptor predated the discovery that hepatoviruses undergo nonlytic release from infected cells as membrane-cloaked, quasi-enveloped HAV (eHAV) virions that enter cells via a pathway distinct from naked, nonenveloped virions. We thus revisited the role of TIM1 in hepatovirus entry, examining both adherence and infection/replication in cells with clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-engineered TIM1 knockout. Cell culture-derived, gradient-purified eHAV bound Huh-7.5 human hepatoma cells less efficiently than naked HAV at 4°C, but eliminating TIM1 expression caused no difference in adherence of either form of HAV, nor any impact on infection and replication in these cells. In contrast, TIM1-deficient Vero cells showed a modest reduction in quasi-enveloped eHAV (but not naked HAV) attachment and replication. Thus, TIM1 facilitates quasi-enveloped eHAV entry in Vero cells, most likely by binding phosphatidylserine (PtdSer) residues on the eHAV membrane. Both Tim1(−/−) Ifnar1(−/−) and Tim4(−/−) Ifnar1(−/−) double-knockout mice were susceptible to infection upon intravenous challenge with infected liver homogenate, with fecal HAV shedding and serum alanine aminotransferase (ALT) elevations similar to those in Ifnar1(−/−) mice. However, intrahepatic HAV RNA and ALT elevations were modestly reduced in Tim1(−/−)Ifnar1(−/−) mice compared to Ifnar1(−/−) mice challenged with a lower titer of gradient-purified HAV or eHAV. We conclude that TIM1 is not an essential hepatovirus entry factor, although its PtdSer-binding activity may contribute to the spread of quasi-enveloped virus and liver injury in mice.
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spelling pubmed-55879072017-09-13 TIM1 (HAVCR1) Is Not Essential for Cellular Entry of Either Quasi-enveloped or Naked Hepatitis A Virions Das, Anshuman Hirai-Yuki, Asuka González-López, Olga Rhein, Bethany Moller-Tank, Sven Brouillette, Rachel Hensley, Lucinda Misumi, Ichiro Lovell, William Cullen, John M. Whitmire, Jason K. Maury, Wendy Lemon, Stanley M. mBio Research Article Receptor molecules play key roles in the cellular entry of picornaviruses, and TIM1 (HAVCR1) is widely accepted to be the receptor for hepatitis A virus (HAV), an unusual, hepatotropic human picornavirus. However, its identification as the hepatovirus receptor predated the discovery that hepatoviruses undergo nonlytic release from infected cells as membrane-cloaked, quasi-enveloped HAV (eHAV) virions that enter cells via a pathway distinct from naked, nonenveloped virions. We thus revisited the role of TIM1 in hepatovirus entry, examining both adherence and infection/replication in cells with clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-engineered TIM1 knockout. Cell culture-derived, gradient-purified eHAV bound Huh-7.5 human hepatoma cells less efficiently than naked HAV at 4°C, but eliminating TIM1 expression caused no difference in adherence of either form of HAV, nor any impact on infection and replication in these cells. In contrast, TIM1-deficient Vero cells showed a modest reduction in quasi-enveloped eHAV (but not naked HAV) attachment and replication. Thus, TIM1 facilitates quasi-enveloped eHAV entry in Vero cells, most likely by binding phosphatidylserine (PtdSer) residues on the eHAV membrane. Both Tim1(−/−) Ifnar1(−/−) and Tim4(−/−) Ifnar1(−/−) double-knockout mice were susceptible to infection upon intravenous challenge with infected liver homogenate, with fecal HAV shedding and serum alanine aminotransferase (ALT) elevations similar to those in Ifnar1(−/−) mice. However, intrahepatic HAV RNA and ALT elevations were modestly reduced in Tim1(−/−)Ifnar1(−/−) mice compared to Ifnar1(−/−) mice challenged with a lower titer of gradient-purified HAV or eHAV. We conclude that TIM1 is not an essential hepatovirus entry factor, although its PtdSer-binding activity may contribute to the spread of quasi-enveloped virus and liver injury in mice. American Society for Microbiology 2017-09-05 /pmc/articles/PMC5587907/ /pubmed/28874468 http://dx.doi.org/10.1128/mBio.00969-17 Text en Copyright © 2017 Das et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Das, Anshuman
Hirai-Yuki, Asuka
González-López, Olga
Rhein, Bethany
Moller-Tank, Sven
Brouillette, Rachel
Hensley, Lucinda
Misumi, Ichiro
Lovell, William
Cullen, John M.
Whitmire, Jason K.
Maury, Wendy
Lemon, Stanley M.
TIM1 (HAVCR1) Is Not Essential for Cellular Entry of Either Quasi-enveloped or Naked Hepatitis A Virions
title TIM1 (HAVCR1) Is Not Essential for Cellular Entry of Either Quasi-enveloped or Naked Hepatitis A Virions
title_full TIM1 (HAVCR1) Is Not Essential for Cellular Entry of Either Quasi-enveloped or Naked Hepatitis A Virions
title_fullStr TIM1 (HAVCR1) Is Not Essential for Cellular Entry of Either Quasi-enveloped or Naked Hepatitis A Virions
title_full_unstemmed TIM1 (HAVCR1) Is Not Essential for Cellular Entry of Either Quasi-enveloped or Naked Hepatitis A Virions
title_short TIM1 (HAVCR1) Is Not Essential for Cellular Entry of Either Quasi-enveloped or Naked Hepatitis A Virions
title_sort tim1 (havcr1) is not essential for cellular entry of either quasi-enveloped or naked hepatitis a virions
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5587907/
https://www.ncbi.nlm.nih.gov/pubmed/28874468
http://dx.doi.org/10.1128/mBio.00969-17
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