Inactivation of the type II receptor reveals two receptor pathways for the diverse TGF-β activities

Science

Volumn 260, Issue 5112, 1993, Pages 1335-1338

  Chen, Ruey Hwa ^a   Ebner, Reinhard ^a   Derynck, Rik ^a  

^a UNIVERSITY OF CALIFORNIA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

MONOCLONAL ANTIBODY; SIGNAL RECOGNITION PARTICLE; TRANSFORMING GROWTH FACTOR BETA;

ARTICLE; CELL DIFFERENTIATION; CELL PROLIFERATION; HUMAN; PHOSPHORYLATION; PRIORITY JOURNAL;

ANIMAL; CELL DIVISION; CELL LINE; DNA; DOWN-REGULATION; FIBRONECTINS; PLASMINOGEN ACTIVATOR INHIBITOR 1; PROTO-ONCOGENE PROTEINS C-JUN; RECEPTORS, CELL SURFACE; RETINOBLASTOMA PROTEIN; SIGNAL TRANSDUCTION; SUPPORT, NON-U.S. GOV'T; SUPPORT, U.S. GOV'T, P.H.S.; TRANSFECTION; TRANSFORMING GROWTH FACTOR BETA;

EID: 0027250810     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.8388126     Document Type: Article

References (31)

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25. To generate the truncated type II receptor, we made an Eco RI-Hpa I fragment corresponding to nucleotides -7 through 573 of the type II receptor by polymerase chain reaction (PCR), followed by cleavage at the unique Hpa I site located immediately 3′ of the transmembrane domain sequence. A double-stranded oligonucleotide adaptor for the sequence encoding the epitope tag, FLAG (22), was ligated to the 3′ end of the truncated type II cDNA, and the resulting fragment was inserted into the Eco RI and Xba I sites of the expression vector pRK5, thus generating the expression plasmid for the truncated type II receptor. DNA was transfected into the QT6 cells and 293 cells by calcium phosphate precipitation (23). Cross-linking analysis was performed as described (3, 24), and the cross-linked proteins were separated by denaturing and reducing polyacrylamide (7.5%) gel electrophoresis and subsequent autoradiography.
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31. We thank H. Lin and R. Weinberg for providing the type II TGF-β receptor cDNA and E. Filvaroff, Z. Werb, and K. Yamamoto for helpful discussions. Supported by grants from the American Cancer Society and the National Institutes of Health (to R.D.) and by a fellowship from the Deutsche Forschungsgemeinschaft (to R.E.).