Stabilization of TM Trimer Interactions during Activation of Moloney Murine Leukemia Virus Env

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Published inJournal of Virology Vol. 82; no. 5; pp. 2358 - 2366
Main Authors Sjöberg, Mathilda, Lindqvist, Birgitta, Garoff, Henrik
Format Journal Article
LanguageEnglish
Published Washington, DC American Society for Microbiology 01.03.2008
American Society for Microbiology (ASM)
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AbstractList The transmembrane subunit (TM) of the trimeric retrovirus Env complex is thought to direct virus-cell membrane fusion by refolding into a cell membrane-interacting, extended form that subsequently folds back on itself into a very stable trimer of hairpin-like TM polypeptides. However, so far there is only limited evidence for the formation of a stable TM trimer during Env activation. Here we have studied the oligomer composition and stability of an intermediate and the fully activated form of Moloney murine leukemia virus (Mo-MLV) Env. Activation of Mo-MLV Env is controlled by isomerization of its intersubunit disulfide. This results in surface subunit (SU) dissociation and TM refolding. If activation is done in the presence of an alkylator, this will modify the isomerization-active thiol in the SU of Env and arrest Env at an intermediate stage, the isomerization-arrested state (IAS) of its activation pathway. We generated IAS and fully activated Envs in vitro and in vivo and studied their states of oligomerization by two-dimensional blue native polyacrylamide gel electrophoresis (PAGE) and nonreducing sodium dodecyl sulfate (SDS)-PAGE. The IAS Env was composed of trimers of SU-TM complexes, whereas the activated Env consisted of SU monomers and TM trimers. When the oligomers were subjected to mild SDS treatment the TM trimer was found to be 3.5 times more resistant than the IAS oligomer. Thus, this demonstrates that a structural conversion of TM takes place during activation, which results in the formation of a stable TM trimer.
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ABSTRACT The transmembrane subunit (TM) of the trimeric retrovirus Env complex is thought to direct virus-cell membrane fusion by refolding into a cell membrane-interacting, extended form that subsequently folds back on itself into a very stable trimer of hairpin-like TM polypeptides. However, so far there is only limited evidence for the formation of a stable TM trimer during Env activation. Here we have studied the oligomer composition and stability of an intermediate and the fully activated form of Moloney murine leukemia virus (Mo-MLV) Env. Activation of Mo-MLV Env is controlled by isomerization of its intersubunit disulfide. This results in surface subunit (SU) dissociation and TM refolding. If activation is done in the presence of an alkylator, this will modify the isomerization-active thiol in the SU of Env and arrest Env at an intermediate stage, the isomerization-arrested state (IAS) of its activation pathway. We generated IAS and fully activated Envs in vitro and in vivo and studied their states of oligomerization by two-dimensional blue native polyacrylamide gel electrophoresis (PAGE) and nonreducing sodium dodecyl sulfate (SDS)-PAGE. The IAS Env was composed of trimers of SU-TM complexes, whereas the activated Env consisted of SU monomers and TM trimers. When the oligomers were subjected to mild SDS treatment the TM trimer was found to be 3.5 times more resistant than the IAS oligomer. Thus, this demonstrates that a structural conversion of TM takes place during activation, which results in the formation of a stable TM trimer.
Author Henrik Garoff
Birgitta Lindqvist
Mathilda Sjöberg
AuthorAffiliation Department of Biosciences and Nutrition, Karolinska Institute, S-141 57 Huddinge, Sweden
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Keywords Virus
Retroviridae
Gammaretrovirus
Interaction
Moloney murine leukemia virus
Virology
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Corresponding author. Mailing address: Department of Biosciences and Nutrition, Karolinska Institute, S-141 57 Huddinge, Sweden. Phone: 46-8-6089125. Fax: 46-8-7745538. E-mail: henrik.garoff@cbt.ki.se
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The transmembrane subunit (TM) of the trimeric retrovirus Env complex is thought to direct virus-cell membrane fusion by refolding into a cell...
ABSTRACT The transmembrane subunit (TM) of the trimeric retrovirus Env complex is thought to direct virus-cell membrane fusion by refolding into a cell...
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SubjectTerms Biological and medical sciences
Biopolymers - metabolism
Cell Line
Disulfides - metabolism
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Gene Products, env - chemistry
Gene Products, env - metabolism
Isomerism
Medicin och hälsovetenskap
Microbiology
Miscellaneous
Moloney murine leukemia virus
Moloney murine leukemia virus - metabolism
Protein Conformation
Protein Folding
Retrovirus
Virology
Virus-Cell Interactions
Title Stabilization of TM Trimer Interactions during Activation of Moloney Murine Leukemia Virus Env
URI http://jvi.asm.org/content/82/5/2358.abstract
https://www.ncbi.nlm.nih.gov/pubmed/18094169
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