Abstract
The cT mutation of PARA, an SV40-adenovirus 7 hybrid virus, abolishes the transport of SV40 T-antigen (T-ag) to the nucleus and results in the accumulation of T-ag in the cytoplasm of infected and transformed cells. The effect of the cT mutation on the association of T-ag with the cell surface was examined in PARA(cT)-infected monkey cells. Surface-associated T-ag was detected by immunofluorescence,lactoperoxidase-catalyzed cell-surface iodination, and cellular fractionation. Surface-associated T-ag was more readily labeled by cell-surface iodination with PARA(cT)-infected cells than with SV40 and wild-type PARA [PARA(nT)]-infected cells. Control experiments eliminated the possibility that the enhanced detection of T-ag in PARA(cT)-infected cells was due to the labeling of intracellular T-ag by lactoperoxidase or to T-ag leaking from dead cells and adhering to the cell surface. The elevated level of surface-associated T-ag in PARA(cT)-infected cells was confirmed by metabolic labeling and cellular fractionation, indicating that the increased iodination of surface-associated T-ag was due to a greater number of T-ag polypeptides on the surface rather than a greater exposure of tyrosine residues. Pulse-chase experiments demonstrated that the elevated level of surface-associated T-ag in PARA(cT)-infected cells was not due to a prolonged stability of surface-associated T-ag. The cT mutation provides a useful tool for studies of mechanisms governing intracellular protein transport.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 169-184 |
| Number of pages | 16 |
| Journal | Virology |
| Volume | 119 |
| Issue number | 1 |
| DOIs | |
| State | Published - 1982 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Virology
- Infectious Diseases
Cite this
Intracellular transport of SV40 large tumor antigen : A mutation which abolishes migration to the nucleus does not prevent association with the cell surface. / Lanford, Robert E.; Butel, Janet S.
In: Virology, Vol. 119, No. 1, 1982, p. 169-184.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Intracellular transport of SV40 large tumor antigen
T2 - A mutation which abolishes migration to the nucleus does not prevent association with the cell surface
AU - Lanford, Robert E.
AU - Butel, Janet S.
PY - 1982
Y1 - 1982
N2 - The cT mutation of PARA, an SV40-adenovirus 7 hybrid virus, abolishes the transport of SV40 T-antigen (T-ag) to the nucleus and results in the accumulation of T-ag in the cytoplasm of infected and transformed cells. The effect of the cT mutation on the association of T-ag with the cell surface was examined in PARA(cT)-infected monkey cells. Surface-associated T-ag was detected by immunofluorescence,lactoperoxidase-catalyzed cell-surface iodination, and cellular fractionation. Surface-associated T-ag was more readily labeled by cell-surface iodination with PARA(cT)-infected cells than with SV40 and wild-type PARA [PARA(nT)]-infected cells. Control experiments eliminated the possibility that the enhanced detection of T-ag in PARA(cT)-infected cells was due to the labeling of intracellular T-ag by lactoperoxidase or to T-ag leaking from dead cells and adhering to the cell surface. The elevated level of surface-associated T-ag in PARA(cT)-infected cells was confirmed by metabolic labeling and cellular fractionation, indicating that the increased iodination of surface-associated T-ag was due to a greater number of T-ag polypeptides on the surface rather than a greater exposure of tyrosine residues. Pulse-chase experiments demonstrated that the elevated level of surface-associated T-ag in PARA(cT)-infected cells was not due to a prolonged stability of surface-associated T-ag. The cT mutation provides a useful tool for studies of mechanisms governing intracellular protein transport.
AB - The cT mutation of PARA, an SV40-adenovirus 7 hybrid virus, abolishes the transport of SV40 T-antigen (T-ag) to the nucleus and results in the accumulation of T-ag in the cytoplasm of infected and transformed cells. The effect of the cT mutation on the association of T-ag with the cell surface was examined in PARA(cT)-infected monkey cells. Surface-associated T-ag was detected by immunofluorescence,lactoperoxidase-catalyzed cell-surface iodination, and cellular fractionation. Surface-associated T-ag was more readily labeled by cell-surface iodination with PARA(cT)-infected cells than with SV40 and wild-type PARA [PARA(nT)]-infected cells. Control experiments eliminated the possibility that the enhanced detection of T-ag in PARA(cT)-infected cells was due to the labeling of intracellular T-ag by lactoperoxidase or to T-ag leaking from dead cells and adhering to the cell surface. The elevated level of surface-associated T-ag in PARA(cT)-infected cells was confirmed by metabolic labeling and cellular fractionation, indicating that the increased iodination of surface-associated T-ag was due to a greater number of T-ag polypeptides on the surface rather than a greater exposure of tyrosine residues. Pulse-chase experiments demonstrated that the elevated level of surface-associated T-ag in PARA(cT)-infected cells was not due to a prolonged stability of surface-associated T-ag. The cT mutation provides a useful tool for studies of mechanisms governing intracellular protein transport.
UR - http://www.scopus.com/inward/record.url?scp=0020033089&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0020033089&partnerID=8YFLogxK
U2 - 10.1016/0042-6822(82)90074-5
DO - 10.1016/0042-6822(82)90074-5
M3 - Article
C2 - 6280381
AN - SCOPUS:0020033089
VL - 119
SP - 169
EP - 184
JO - Virology
JF - Virology
SN - 0042-6822
IS - 1
ER -