Molecular Determinants of Human T-cell Leukemia Virus Type 1 Gag Targeting to the Plasma Membrane for Assembly

Dominik Herrmann; Heather M Hanson; Lynne W. Zhou; Rayna Addabbo; Nora A Willkomm; Isaac Angert; Joachim D. Mueller; Louis M. Mansky; Jamil S. Saad

doi:10.1016/j.jmb.2022.167609

Molecular Determinants of Human T-cell Leukemia Virus Type 1 Gag Targeting to the Plasma Membrane for Assembly

Dominik Herrmann, Heather M Hanson, Lynne W. Zhou, Rayna Addabbo, Nora A Willkomm, Isaac Angert, Joachim D. Mueller, Louis M. Mansky, Jamil S. Saad

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Assembly of human T-cell leukemia virus type 1 (HTLV-1) particles is initiated by the trafficking of virally encoded Gag polyproteins to the inner leaflet of the plasma membrane (PM). Gag–PM interactions are mediated by the matrix (MA) domain, which contains a myristoyl group (myr) and a basic patch formed by lysine and arginine residues. For many retroviruses, Gag–PM interactions are mediated by phosphatidylinositol 4,5-bisphosphate [PI(4,5)P₂]; however, previous studies suggested that HTLV-1 Gag–PM interactions and therefore virus assembly are less dependent on PI(4,5)P₂. We have recently shown that PI(4,5)P₂ binds directly to HTLV-1 unmyristoylated MA [myr(–)MA] and that myr(–)MA binding to membranes is significantly enhanced by inclusion of phosphatidylserine (PS) and PI(4,5)P₂. Herein, we employed structural, biophysical, biochemical, mutagenesis, and cell-based assays to identify residues involved in MA–membrane interactions. Our data revealed that the lysine-rich motif (Lys47, Lys48, and Lys51) constitutes the primary PI(4,5)P₂–binding site. Furthermore, we show that arginine residues 3, 7, 14 and 17 located in the unstructured N-terminus are essential for MA binding to membranes containing PS and/or PI(4,5)P₂. Substitution of lysine and arginine residues severely attenuated virus-like particle production, but only the lysine residues could be clearly correlated with reduced PM binding. These results support a mechanism by which HTLV-1 Gag targeting to the PM is mediated by a trio engagement of the myr group, Arg-rich and Lys-rich motifs. These findings advance our understanding of a key step in retroviral particle assembly.

Original language	English (US)
Article number	167609
Journal	Journal of Molecular Biology
Volume	434
Issue number	12
DOIs	https://doi.org/10.1016/j.jmb.2022.167609
State	Published - Jun 30 2022

Bibliographical note

Funding Information:
This work was supported by grants 9 R01 AI150901-10 from the National Institutes of Health (NIH) to JSS; R01 GM098550 (to LMM); T32 AI083196 and F31 AI147805 (to HMH), T90 DE022732 and F30 DE031829 (to NAW), TL1R002493 (to RA); and T90 DE022732 (to IA). The High-Field NMR facility at the University of Alabama at Birmingham was established through NIH grant 1S10RR026478 and is currently supported through the O'Neal Comprehensive Cancer center (NCI grant P30 CA013148). The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Funding Information:
This work was supported by grants 9 R01 AI150901-10 from the National Institutes of Health (NIH) to JSS; R01 GM098550 (to LMM); T32 AI083196 and F31 AI147805 (to HMH), T90 DE022732 and F30 DE031829 (to NAW), TL1R002493 (to RA); and T90 DE022732 (to IA). The High-Field NMR facility at the University of Alabama at Birmingham was established through NIH grant 1S10RR026478 and is currently supported through the O’Neal Comprehensive Cancer center (NCI grant P30 CA013148).

Publisher Copyright:
© 2022 Elsevier Ltd

Keywords

Gag polyprotein
human T-cell leukemia virus type 1 (HTLV-1)
human immunodeficiency virus type 1 (HIV-1)
matrix (MA) protein
plasma membrane (PM)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Publisher link

10.1016/j.jmb.2022.167609

Find It

Find It at U of M Libraries

Cite this

@article{efff2ec83cc24d5cbd3c2ca03c3286d7,

title = "Molecular Determinants of Human T-cell Leukemia Virus Type 1 Gag Targeting to the Plasma Membrane for Assembly",

abstract = "Assembly of human T-cell leukemia virus type 1 (HTLV-1) particles is initiated by the trafficking of virally encoded Gag polyproteins to the inner leaflet of the plasma membrane (PM). Gag–PM interactions are mediated by the matrix (MA) domain, which contains a myristoyl group (myr) and a basic patch formed by lysine and arginine residues. For many retroviruses, Gag–PM interactions are mediated by phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2]; however, previous studies suggested that HTLV-1 Gag–PM interactions and therefore virus assembly are less dependent on PI(4,5)P2. We have recently shown that PI(4,5)P2 binds directly to HTLV-1 unmyristoylated MA [myr(–)MA] and that myr(–)MA binding to membranes is significantly enhanced by inclusion of phosphatidylserine (PS) and PI(4,5)P2. Herein, we employed structural, biophysical, biochemical, mutagenesis, and cell-based assays to identify residues involved in MA–membrane interactions. Our data revealed that the lysine-rich motif (Lys47, Lys48, and Lys51) constitutes the primary PI(4,5)P2–binding site. Furthermore, we show that arginine residues 3, 7, 14 and 17 located in the unstructured N-terminus are essential for MA binding to membranes containing PS and/or PI(4,5)P2. Substitution of lysine and arginine residues severely attenuated virus-like particle production, but only the lysine residues could be clearly correlated with reduced PM binding. These results support a mechanism by which HTLV-1 Gag targeting to the PM is mediated by a trio engagement of the myr group, Arg-rich and Lys-rich motifs. These findings advance our understanding of a key step in retroviral particle assembly.",

keywords = "Gag polyprotein, human T-cell leukemia virus type 1 (HTLV-1), human immunodeficiency virus type 1 (HIV-1), matrix (MA) protein, plasma membrane (PM)",

author = "Dominik Herrmann and Hanson, {Heather M} and Zhou, {Lynne W.} and Rayna Addabbo and Willkomm, {Nora A} and Isaac Angert and Mueller, {Joachim D.} and Mansky, {Louis M.} and Saad, {Jamil S.}",

note = "Funding Information: This work was supported by grants 9 R01 AI150901-10 from the National Institutes of Health (NIH) to JSS; R01 GM098550 (to LMM); T32 AI083196 and F31 AI147805 (to HMH), T90 DE022732 and F30 DE031829 (to NAW), TL1R002493 (to RA); and T90 DE022732 (to IA). The High-Field NMR facility at the University of Alabama at Birmingham was established through NIH grant 1S10RR026478 and is currently supported through the O'Neal Comprehensive Cancer center (NCI grant P30 CA013148). The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Funding Information: This work was supported by grants 9 R01 AI150901-10 from the National Institutes of Health (NIH) to JSS; R01 GM098550 (to LMM); T32 AI083196 and F31 AI147805 (to HMH), T90 DE022732 and F30 DE031829 (to NAW), TL1R002493 (to RA); and T90 DE022732 (to IA). The High-Field NMR facility at the University of Alabama at Birmingham was established through NIH grant 1S10RR026478 and is currently supported through the O{\textquoteright}Neal Comprehensive Cancer center (NCI grant P30 CA013148). Publisher Copyright: {\textcopyright} 2022 Elsevier Ltd",

year = "2022",

month = jun,

day = "30",

doi = "10.1016/j.jmb.2022.167609",

language = "English (US)",

volume = "434",

journal = "Journal of Molecular Biology",

issn = "0022-2836",

publisher = "Academic Press Inc.",

number = "12",

}

TY - JOUR

T1 - Molecular Determinants of Human T-cell Leukemia Virus Type 1 Gag Targeting to the Plasma Membrane for Assembly

AU - Herrmann, Dominik

AU - Hanson, Heather M

AU - Zhou, Lynne W.

AU - Addabbo, Rayna

AU - Willkomm, Nora A

AU - Angert, Isaac

AU - Mueller, Joachim D.

AU - Mansky, Louis M.

AU - Saad, Jamil S.

N1 - Funding Information: This work was supported by grants 9 R01 AI150901-10 from the National Institutes of Health (NIH) to JSS; R01 GM098550 (to LMM); T32 AI083196 and F31 AI147805 (to HMH), T90 DE022732 and F30 DE031829 (to NAW), TL1R002493 (to RA); and T90 DE022732 (to IA). The High-Field NMR facility at the University of Alabama at Birmingham was established through NIH grant 1S10RR026478 and is currently supported through the O'Neal Comprehensive Cancer center (NCI grant P30 CA013148). The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Funding Information: This work was supported by grants 9 R01 AI150901-10 from the National Institutes of Health (NIH) to JSS; R01 GM098550 (to LMM); T32 AI083196 and F31 AI147805 (to HMH), T90 DE022732 and F30 DE031829 (to NAW), TL1R002493 (to RA); and T90 DE022732 (to IA). The High-Field NMR facility at the University of Alabama at Birmingham was established through NIH grant 1S10RR026478 and is currently supported through the O’Neal Comprehensive Cancer center (NCI grant P30 CA013148). Publisher Copyright: © 2022 Elsevier Ltd

PY - 2022/6/30

Y1 - 2022/6/30

N2 - Assembly of human T-cell leukemia virus type 1 (HTLV-1) particles is initiated by the trafficking of virally encoded Gag polyproteins to the inner leaflet of the plasma membrane (PM). Gag–PM interactions are mediated by the matrix (MA) domain, which contains a myristoyl group (myr) and a basic patch formed by lysine and arginine residues. For many retroviruses, Gag–PM interactions are mediated by phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2]; however, previous studies suggested that HTLV-1 Gag–PM interactions and therefore virus assembly are less dependent on PI(4,5)P2. We have recently shown that PI(4,5)P2 binds directly to HTLV-1 unmyristoylated MA [myr(–)MA] and that myr(–)MA binding to membranes is significantly enhanced by inclusion of phosphatidylserine (PS) and PI(4,5)P2. Herein, we employed structural, biophysical, biochemical, mutagenesis, and cell-based assays to identify residues involved in MA–membrane interactions. Our data revealed that the lysine-rich motif (Lys47, Lys48, and Lys51) constitutes the primary PI(4,5)P2–binding site. Furthermore, we show that arginine residues 3, 7, 14 and 17 located in the unstructured N-terminus are essential for MA binding to membranes containing PS and/or PI(4,5)P2. Substitution of lysine and arginine residues severely attenuated virus-like particle production, but only the lysine residues could be clearly correlated with reduced PM binding. These results support a mechanism by which HTLV-1 Gag targeting to the PM is mediated by a trio engagement of the myr group, Arg-rich and Lys-rich motifs. These findings advance our understanding of a key step in retroviral particle assembly.

AB - Assembly of human T-cell leukemia virus type 1 (HTLV-1) particles is initiated by the trafficking of virally encoded Gag polyproteins to the inner leaflet of the plasma membrane (PM). Gag–PM interactions are mediated by the matrix (MA) domain, which contains a myristoyl group (myr) and a basic patch formed by lysine and arginine residues. For many retroviruses, Gag–PM interactions are mediated by phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2]; however, previous studies suggested that HTLV-1 Gag–PM interactions and therefore virus assembly are less dependent on PI(4,5)P2. We have recently shown that PI(4,5)P2 binds directly to HTLV-1 unmyristoylated MA [myr(–)MA] and that myr(–)MA binding to membranes is significantly enhanced by inclusion of phosphatidylserine (PS) and PI(4,5)P2. Herein, we employed structural, biophysical, biochemical, mutagenesis, and cell-based assays to identify residues involved in MA–membrane interactions. Our data revealed that the lysine-rich motif (Lys47, Lys48, and Lys51) constitutes the primary PI(4,5)P2–binding site. Furthermore, we show that arginine residues 3, 7, 14 and 17 located in the unstructured N-terminus are essential for MA binding to membranes containing PS and/or PI(4,5)P2. Substitution of lysine and arginine residues severely attenuated virus-like particle production, but only the lysine residues could be clearly correlated with reduced PM binding. These results support a mechanism by which HTLV-1 Gag targeting to the PM is mediated by a trio engagement of the myr group, Arg-rich and Lys-rich motifs. These findings advance our understanding of a key step in retroviral particle assembly.

KW - Gag polyprotein

KW - human T-cell leukemia virus type 1 (HTLV-1)

KW - human immunodeficiency virus type 1 (HIV-1)

KW - matrix (MA) protein

KW - plasma membrane (PM)

UR - http://www.scopus.com/inward/record.url?scp=85130378175&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85130378175&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2022.167609

DO - 10.1016/j.jmb.2022.167609

M3 - Article

C2 - 35490898

AN - SCOPUS:85130378175

SN - 0022-2836

VL - 434

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 12

M1 - 167609

ER -

Molecular Determinants of Human T-cell Leukemia Virus Type 1 Gag Targeting to the Plasma Membrane for Assembly

Abstract

Bibliographical note

Keywords

UN SDGs

Publisher link

Find It

Other files and links

Fingerprint

Cite this