Retroviral envelope protein fusions to secreted and membrane markers

M. Catherine Mace, Mark Hansen, Sam Whiting, Chin Tien Wang, Eric Barklis

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We have analyzed a series of Moloney murine leukemia (M-MuLV) envelope (env) protein fusions to the marker proteins invertase and placental alkaline phosphatase (PLAP), expressed in Psi2 retrovirus packaging cells. he yeast invertase protein, fused at its third amino acid residue to the amino-terminal signal sequence and 17 residues of the mature M-MuLV env protein, retained its enzymatic activity and was secreted from mammalian cells. However, env protein fusions to the C-terminal portion of invertase were inactive. In contrast, some, but not all, env protein fusions at the C-terminal region of PLAP were enzymatically active: PLAP fusions containing long C-terminal portions of env localized to the rough endoplasmic reticulum (RER) and possessed low enzyme activity levels, while fusion constructs containing relatively short portions of the M-MuLV env gene localized to the Golgi and had higher activity levels. Those proteins that localized to the Golgi also were processed, in part, to forms of 67 to 68 kDa, the size of the mature PLAP protein. Since PLAP ordinarily is transfered to a phosphatidyl-inositol glycan tail (PIG-tail) in the Golgi and then transported to the plasma membrane, it appears that Golgi-localized PLAP-env fusions are processed imperfectly. PLAP itself, when expressed in Psi2 cells, accumulated at the plasma membrane and, unlike the PIG-tailed Thy-1 protein, was not incorporated into virus particles. Thus, the reported incorporation of the Thy-1 protein into M-MuLV virions does not appear to be a consequence of its glycoprotein tail.

Original languageEnglish (US)
Pages (from-to)869-874
Number of pages6
JournalVirology
Volume188
Issue number2
DOIs
StatePublished - 1992

Fingerprint

Membranes
beta-Fructofuranosidase
Proteins
Virion
Cell Membrane
Fungal Proteins
Rough Endoplasmic Reticulum
Product Packaging
Retroviridae
Protein Sorting Signals
Phosphatidylinositols
placental alkaline phosphatase
Glycoproteins
Leukemia
Amino Acids
Enzymes
Genes

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Retroviral envelope protein fusions to secreted and membrane markers. / Mace, M. Catherine; Hansen, Mark; Whiting, Sam; Wang, Chin Tien; Barklis, Eric.

In: Virology, Vol. 188, No. 2, 1992, p. 869-874.

Research output: Contribution to journalArticle

Mace, M. Catherine ; Hansen, Mark ; Whiting, Sam ; Wang, Chin Tien ; Barklis, Eric. / Retroviral envelope protein fusions to secreted and membrane markers. In: Virology. 1992 ; Vol. 188, No. 2. pp. 869-874.
@article{2f1f8847ad8f4233a0b180910951aeed,
title = "Retroviral envelope protein fusions to secreted and membrane markers",
abstract = "We have analyzed a series of Moloney murine leukemia (M-MuLV) envelope (env) protein fusions to the marker proteins invertase and placental alkaline phosphatase (PLAP), expressed in Psi2 retrovirus packaging cells. he yeast invertase protein, fused at its third amino acid residue to the amino-terminal signal sequence and 17 residues of the mature M-MuLV env protein, retained its enzymatic activity and was secreted from mammalian cells. However, env protein fusions to the C-terminal portion of invertase were inactive. In contrast, some, but not all, env protein fusions at the C-terminal region of PLAP were enzymatically active: PLAP fusions containing long C-terminal portions of env localized to the rough endoplasmic reticulum (RER) and possessed low enzyme activity levels, while fusion constructs containing relatively short portions of the M-MuLV env gene localized to the Golgi and had higher activity levels. Those proteins that localized to the Golgi also were processed, in part, to forms of 67 to 68 kDa, the size of the mature PLAP protein. Since PLAP ordinarily is transfered to a phosphatidyl-inositol glycan tail (PIG-tail) in the Golgi and then transported to the plasma membrane, it appears that Golgi-localized PLAP-env fusions are processed imperfectly. PLAP itself, when expressed in Psi2 cells, accumulated at the plasma membrane and, unlike the PIG-tailed Thy-1 protein, was not incorporated into virus particles. Thus, the reported incorporation of the Thy-1 protein into M-MuLV virions does not appear to be a consequence of its glycoprotein tail.",
author = "Mace, {M. Catherine} and Mark Hansen and Sam Whiting and Wang, {Chin Tien} and Eric Barklis",
year = "1992",
doi = "10.1016/0042-6822(92)90544-Y",
language = "English (US)",
volume = "188",
pages = "869--874",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Retroviral envelope protein fusions to secreted and membrane markers

AU - Mace, M. Catherine

AU - Hansen, Mark

AU - Whiting, Sam

AU - Wang, Chin Tien

AU - Barklis, Eric

PY - 1992

Y1 - 1992

N2 - We have analyzed a series of Moloney murine leukemia (M-MuLV) envelope (env) protein fusions to the marker proteins invertase and placental alkaline phosphatase (PLAP), expressed in Psi2 retrovirus packaging cells. he yeast invertase protein, fused at its third amino acid residue to the amino-terminal signal sequence and 17 residues of the mature M-MuLV env protein, retained its enzymatic activity and was secreted from mammalian cells. However, env protein fusions to the C-terminal portion of invertase were inactive. In contrast, some, but not all, env protein fusions at the C-terminal region of PLAP were enzymatically active: PLAP fusions containing long C-terminal portions of env localized to the rough endoplasmic reticulum (RER) and possessed low enzyme activity levels, while fusion constructs containing relatively short portions of the M-MuLV env gene localized to the Golgi and had higher activity levels. Those proteins that localized to the Golgi also were processed, in part, to forms of 67 to 68 kDa, the size of the mature PLAP protein. Since PLAP ordinarily is transfered to a phosphatidyl-inositol glycan tail (PIG-tail) in the Golgi and then transported to the plasma membrane, it appears that Golgi-localized PLAP-env fusions are processed imperfectly. PLAP itself, when expressed in Psi2 cells, accumulated at the plasma membrane and, unlike the PIG-tailed Thy-1 protein, was not incorporated into virus particles. Thus, the reported incorporation of the Thy-1 protein into M-MuLV virions does not appear to be a consequence of its glycoprotein tail.

AB - We have analyzed a series of Moloney murine leukemia (M-MuLV) envelope (env) protein fusions to the marker proteins invertase and placental alkaline phosphatase (PLAP), expressed in Psi2 retrovirus packaging cells. he yeast invertase protein, fused at its third amino acid residue to the amino-terminal signal sequence and 17 residues of the mature M-MuLV env protein, retained its enzymatic activity and was secreted from mammalian cells. However, env protein fusions to the C-terminal portion of invertase were inactive. In contrast, some, but not all, env protein fusions at the C-terminal region of PLAP were enzymatically active: PLAP fusions containing long C-terminal portions of env localized to the rough endoplasmic reticulum (RER) and possessed low enzyme activity levels, while fusion constructs containing relatively short portions of the M-MuLV env gene localized to the Golgi and had higher activity levels. Those proteins that localized to the Golgi also were processed, in part, to forms of 67 to 68 kDa, the size of the mature PLAP protein. Since PLAP ordinarily is transfered to a phosphatidyl-inositol glycan tail (PIG-tail) in the Golgi and then transported to the plasma membrane, it appears that Golgi-localized PLAP-env fusions are processed imperfectly. PLAP itself, when expressed in Psi2 cells, accumulated at the plasma membrane and, unlike the PIG-tailed Thy-1 protein, was not incorporated into virus particles. Thus, the reported incorporation of the Thy-1 protein into M-MuLV virions does not appear to be a consequence of its glycoprotein tail.

UR - http://www.scopus.com/inward/record.url?scp=0026690971&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026690971&partnerID=8YFLogxK

U2 - 10.1016/0042-6822(92)90544-Y

DO - 10.1016/0042-6822(92)90544-Y

M3 - Article

C2 - 1585654

AN - SCOPUS:0026690971

VL - 188

SP - 869

EP - 874

JO - Virology

JF - Virology

SN - 0042-6822

IS - 2

ER -