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Nucleic Acids Research, 1983, Vol. 11, No. 18 6199-6210
© 1983

MOLECULAR BIOLOGY

Isolation of cDNA clones for basal lamina components: type IV procollagen

Markku Kurkinen, Denise P Barlow, David M. Helfman*, Jeffrey G. Williams and Brigid L.M. Hogan

Imperial Cancer Research Fund, Mill Hill Laboratories Burtonhole Lane, London NW7 1AD, UK *Cold Spring Harbor Laboratory P.O. Box 100, Cold Spring Harbor, NY 11724, USA

Received July 28, 1983. Accepted September 6, 1983.

We have isolated cONA clones for mouse type IV procollagen from a library constructed from total poly A+RNA of 13.5 day mouse embryo parietal endoderm (PE) cells. In Northern analysis these clones hybridise to a 6.8 kb RNA which is abundant in embryonic PE cells and in differentiated F9 teratocarcinoma cells. Hybrid selection and in vitro translation of the cDNA specific mRNA produced a single polypeptide of Mr = 165 000. This polypeptide was specifically immunoprecipitated with mouse type IV procollagen antisera and comigrated on SDS-gel electrophoresis with one of the two in vitro synthesised chains of type IV procollagen. Undifferentiated F9 teratocarcinoma cells can be induced by retinolc acid and dibutyryl cAMP to differentiate in vitro into endodermlike cells which resemble mouse PE cells in syntheslsing large amounts of basement membrane proteins, including type IV procollagen. Here we show, using one of the cDNA clones as a probe for type IV procollagen, that an increase in cellular concentration of type IV procollagen mRNA occurs within 24 to 48 hours of induction, reaching a constant high level by 72 hours.


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