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PMID: 2830486 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Signal-dependent translocation of simian virus 40 large-T antigen into rat liver nuclei in a cell-free system.

Molecular and cellular biology ·Vol. 7 ·No. 12 ·1987-12-00 ·Pages 4255-65

Markland W, Smith AE, Roberts BL

Abstract

An in vitro nuclear translocation system is described in which isolated rat liver nuclei were incubated in a defined buffered medium containing radiolabeled or fluorescently labeled exogenous proteins. The nuclei were rapidly recovered, extracted, and analyzed for the presence of associated radiolabeled or fluorescently labeled proteins. The isolated nuclei exhibited the same specificity for protein uptake as seen previously in vivo, accumulating simian virus 40 wild-type large-T antigen and p53 while excluding a cytoplasmic variant of large-T antigen (d10) and bovine serum albumin. The rapid nuclear accumulation of wild-type large-T antigen was shown to be selective and dependent upon the recognition of a wild-type nuclear location signal, ATP and temperature dependent, and unidirectional. Taken together, the data suggest that in our in vitro system the nuclear translocation of wild-type large-T antigen exhibits some of the characteristics of an active transport process.

MeSH Terms
Animals Antigens, Polyomavirus Transforming/isolation & purification Biological Transport, Active Cell Nucleus/metabolism Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique Fluorescent Dyes Immunosorbent Techniques Liver/ultrastructure Male Mice Molecular Weight Protein Sorting Signals/physiology Pyruvate Kinase/metabolism Rats Rats, Inbred Strains Recombinant Fusion Proteins/metabolism Rhodamines Simian virus 40/immunology Sulfur Radioisotopes
Chemicals
Antigens, Polyomavirus Transforming Fluorescent Dyes Protein Sorting Signals Recombinant Fusion Proteins Rhodamines Sulfur Radioisotopes rhodamine isothiocyanate Pyruvate Kinase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Markland W
Integrated Genetics, Framingham, Massachusetts 01701.
Smith A E
Roberts B L
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36 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.7.12.4255-4265.1987
Published
1987-12-00
Pages
4255-65
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC368107
Subset
IM
Grants
NCI NIH HHS · R01 CA 43186-01 · United States
External Links
PubMed source DOI: 10.1128/mcb.7.12.4255-4265.1987 PMC Full Text View journal details
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