In vitro and in vivo functional analysis of human T cell lymphotropic virus type 1 pX open reading frames I and II

Michael Dale Lairmore, B. Albrecht, C. D'Souza, J. W. Nisbet, W. Ding, J. T. Bartoe, P. L. Green, W. Zhang

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Human T lymphotropic virus type 1 (HTLV-1) is a complex retrovirus containing regulatory and accessory genes encoded in four open reading frames (ORF I-IV) of the pX region. It is not clear what role pX ORFs I and II-encoded proteins have in the pathogenesis of the lymphoproliferative diseases associated with HTLV-1 infection. The conserved ORF I encodes for a hydrophobic 12-kDa protein, p12,(I) that contains four SH3 binding motifs (PXXP) that localizes to cellular endomembranes when overexpressed in cultured cells. Differential splicing of pX ORF II results in the production of two nuclear proteins, p13(II) and p30(II). p13(II) also localizes to mitochondria, p30(II) shares homology with the POU family of transcription factors. We have identified functional roles of pX ORF I and ORF II in establishment and maintenance of infection in a rabbit model. To functionally study p12(I) we have tested a proviral clone with selective ablation of ORF I (ACH.p12(I)) for its ability to infect quiescent peripheral blood mononuclear cells (PBMC). Our data indicate that T cells infected with the wild-type clone of HTLV-1 (ACH) are more efficient than ACH.p12(I) in infecting quiescent PBMC. These findings parallel our animal model data and suggest a role for p12(I) in the activation of quiescent lymphocytes, a prerequisite for effective viral replication in vivo. To test the ability of p30(II) to function as a transcription factor we have constructed p30(II) as a Gal4-fusion protein. When transfected with Gal4-driven luciferase reporter genes, the p30(II)-Gal4-fusion protein induces transcriptional activity up to 50-fold in both 293 and HeLa-Tat cells. These systems will be useful to identify molecular mechanisms that explain the functional role of pX ORF I and ORF II-encoded proteins in HTLV-1 replication.

Original languageEnglish (US)
Pages (from-to)1757-1764
Number of pages8
JournalAIDS Research and Human Retroviruses
Volume16
Issue number16
DOIs
StatePublished - Nov 1 2000
Externally publishedYes

Fingerprint

Human T-lymphotropic virus 1
Open Reading Frames
T-Lymphocytes
Proteins
Blood Cells
Clone Cells
POU Domain Factors
In Vitro Techniques
Virus Diseases
Retroviridae
Regulator Genes
Lymphocyte Activation
Virus Replication
Nuclear Proteins
Luciferases
Reporter Genes
HeLa Cells
Cultured Cells
Mitochondria
Transcription Factors

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

In vitro and in vivo functional analysis of human T cell lymphotropic virus type 1 pX open reading frames I and II. / Lairmore, Michael Dale; Albrecht, B.; D'Souza, C.; Nisbet, J. W.; Ding, W.; Bartoe, J. T.; Green, P. L.; Zhang, W.

In: AIDS Research and Human Retroviruses, Vol. 16, No. 16, 01.11.2000, p. 1757-1764.

Research output: Contribution to journalArticle

Lairmore, Michael Dale ; Albrecht, B. ; D'Souza, C. ; Nisbet, J. W. ; Ding, W. ; Bartoe, J. T. ; Green, P. L. ; Zhang, W. / In vitro and in vivo functional analysis of human T cell lymphotropic virus type 1 pX open reading frames I and II. In: AIDS Research and Human Retroviruses. 2000 ; Vol. 16, No. 16. pp. 1757-1764.
@article{a6ecea7967e6429d9571091a5273612a,
title = "In vitro and in vivo functional analysis of human T cell lymphotropic virus type 1 pX open reading frames I and II",
abstract = "Human T lymphotropic virus type 1 (HTLV-1) is a complex retrovirus containing regulatory and accessory genes encoded in four open reading frames (ORF I-IV) of the pX region. It is not clear what role pX ORFs I and II-encoded proteins have in the pathogenesis of the lymphoproliferative diseases associated with HTLV-1 infection. The conserved ORF I encodes for a hydrophobic 12-kDa protein, p12,(I) that contains four SH3 binding motifs (PXXP) that localizes to cellular endomembranes when overexpressed in cultured cells. Differential splicing of pX ORF II results in the production of two nuclear proteins, p13(II) and p30(II). p13(II) also localizes to mitochondria, p30(II) shares homology with the POU family of transcription factors. We have identified functional roles of pX ORF I and ORF II in establishment and maintenance of infection in a rabbit model. To functionally study p12(I) we have tested a proviral clone with selective ablation of ORF I (ACH.p12(I)) for its ability to infect quiescent peripheral blood mononuclear cells (PBMC). Our data indicate that T cells infected with the wild-type clone of HTLV-1 (ACH) are more efficient than ACH.p12(I) in infecting quiescent PBMC. These findings parallel our animal model data and suggest a role for p12(I) in the activation of quiescent lymphocytes, a prerequisite for effective viral replication in vivo. To test the ability of p30(II) to function as a transcription factor we have constructed p30(II) as a Gal4-fusion protein. When transfected with Gal4-driven luciferase reporter genes, the p30(II)-Gal4-fusion protein induces transcriptional activity up to 50-fold in both 293 and HeLa-Tat cells. These systems will be useful to identify molecular mechanisms that explain the functional role of pX ORF I and ORF II-encoded proteins in HTLV-1 replication.",
author = "Lairmore, {Michael Dale} and B. Albrecht and C. D'Souza and Nisbet, {J. W.} and W. Ding and Bartoe, {J. T.} and Green, {P. L.} and W. Zhang",
year = "2000",
month = "11",
day = "1",
doi = "10.1089/08892220050193272",
language = "English (US)",
volume = "16",
pages = "1757--1764",
journal = "AIDS Research and Human Retroviruses",
issn = "0889-2229",
publisher = "Mary Ann Liebert Inc.",
number = "16",

}

TY - JOUR

T1 - In vitro and in vivo functional analysis of human T cell lymphotropic virus type 1 pX open reading frames I and II

AU - Lairmore, Michael Dale

AU - Albrecht, B.

AU - D'Souza, C.

AU - Nisbet, J. W.

AU - Ding, W.

AU - Bartoe, J. T.

AU - Green, P. L.

AU - Zhang, W.

PY - 2000/11/1

Y1 - 2000/11/1

N2 - Human T lymphotropic virus type 1 (HTLV-1) is a complex retrovirus containing regulatory and accessory genes encoded in four open reading frames (ORF I-IV) of the pX region. It is not clear what role pX ORFs I and II-encoded proteins have in the pathogenesis of the lymphoproliferative diseases associated with HTLV-1 infection. The conserved ORF I encodes for a hydrophobic 12-kDa protein, p12,(I) that contains four SH3 binding motifs (PXXP) that localizes to cellular endomembranes when overexpressed in cultured cells. Differential splicing of pX ORF II results in the production of two nuclear proteins, p13(II) and p30(II). p13(II) also localizes to mitochondria, p30(II) shares homology with the POU family of transcription factors. We have identified functional roles of pX ORF I and ORF II in establishment and maintenance of infection in a rabbit model. To functionally study p12(I) we have tested a proviral clone with selective ablation of ORF I (ACH.p12(I)) for its ability to infect quiescent peripheral blood mononuclear cells (PBMC). Our data indicate that T cells infected with the wild-type clone of HTLV-1 (ACH) are more efficient than ACH.p12(I) in infecting quiescent PBMC. These findings parallel our animal model data and suggest a role for p12(I) in the activation of quiescent lymphocytes, a prerequisite for effective viral replication in vivo. To test the ability of p30(II) to function as a transcription factor we have constructed p30(II) as a Gal4-fusion protein. When transfected with Gal4-driven luciferase reporter genes, the p30(II)-Gal4-fusion protein induces transcriptional activity up to 50-fold in both 293 and HeLa-Tat cells. These systems will be useful to identify molecular mechanisms that explain the functional role of pX ORF I and ORF II-encoded proteins in HTLV-1 replication.

AB - Human T lymphotropic virus type 1 (HTLV-1) is a complex retrovirus containing regulatory and accessory genes encoded in four open reading frames (ORF I-IV) of the pX region. It is not clear what role pX ORFs I and II-encoded proteins have in the pathogenesis of the lymphoproliferative diseases associated with HTLV-1 infection. The conserved ORF I encodes for a hydrophobic 12-kDa protein, p12,(I) that contains four SH3 binding motifs (PXXP) that localizes to cellular endomembranes when overexpressed in cultured cells. Differential splicing of pX ORF II results in the production of two nuclear proteins, p13(II) and p30(II). p13(II) also localizes to mitochondria, p30(II) shares homology with the POU family of transcription factors. We have identified functional roles of pX ORF I and ORF II in establishment and maintenance of infection in a rabbit model. To functionally study p12(I) we have tested a proviral clone with selective ablation of ORF I (ACH.p12(I)) for its ability to infect quiescent peripheral blood mononuclear cells (PBMC). Our data indicate that T cells infected with the wild-type clone of HTLV-1 (ACH) are more efficient than ACH.p12(I) in infecting quiescent PBMC. These findings parallel our animal model data and suggest a role for p12(I) in the activation of quiescent lymphocytes, a prerequisite for effective viral replication in vivo. To test the ability of p30(II) to function as a transcription factor we have constructed p30(II) as a Gal4-fusion protein. When transfected with Gal4-driven luciferase reporter genes, the p30(II)-Gal4-fusion protein induces transcriptional activity up to 50-fold in both 293 and HeLa-Tat cells. These systems will be useful to identify molecular mechanisms that explain the functional role of pX ORF I and ORF II-encoded proteins in HTLV-1 replication.

UR - http://www.scopus.com/inward/record.url?scp=0034331210&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034331210&partnerID=8YFLogxK

U2 - 10.1089/08892220050193272

DO - 10.1089/08892220050193272

M3 - Article

C2 - 11080823

AN - SCOPUS:0034331210

VL - 16

SP - 1757

EP - 1764

JO - AIDS Research and Human Retroviruses

JF - AIDS Research and Human Retroviruses

SN - 0889-2229

IS - 16

ER -