Clathrin-coated pits with long, dynamin-wrapped necks upon expression of a clathrin antisense RNA
- T.-G. Iversen*,†,
- G. Skretting*,†,‡,
- B. van Deurs§, and
- K. Sandvig*,¶
- *Institute for Cancer Research at the Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway; and §Structural Cell Biology Unit, Department of Anatomy, The Panum Institute, University of Copenhagen, DK-2200 Copenhagen N, Denmark
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Edited by Pietro V. De Camilli, Yale University School of Medicine, New Haven, CT, and approved January 14, 2003 (received for review July 15, 2002)
Abstract
To investigate the role of clathrin in coated vesicle formation, a cell line with inducible expression of clathrin heavy chain (CHC) antisense RNA was produced. After 18 h of CHC antisense RNA expression, the internalization of transferrin was inhibited by 90%. Although the amount of CHC was reduced by only 10%, the frequency of clathrin-coated pits at the cell surface increased by a factor of 3–5, and clathrin-coated structures also accumulated on a pleiomorphic, multivesicular, endosomal compartment. Remarkably, the coated pits were connected to the cell surface by long, tubular necks wrapped by dynamin rings, and the level of dynamin in the CHC antisense RNA-expressing cells was up-regulated 10-fold. In contrast, the amount of several other proteins associated with clathrin coat formation was unaffected. Thus, this study demonstrates that CHC antisense RNA causes accumulation of clathrin-coated pits with dynamin rings around the neck in intact cells not transfected with dynamin mutants, suggesting the existence of a previously uncharacterized functional interplay between clathrin and dynamin.
Footnotes
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↵ † T.-G.I. and G.S. contributed equally to this work.
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↵ ‡ Present address: Norwegian School of Veterinary Science, N-0033 Oslo, Norway.
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↵ ¶ To whom correspondence should be addressed. E-mail: ksandvig{at}radium.uio.no.
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This paper was submitted directly (Track II) to the PNAS office.
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See commentary on page 4981.
- Abbreviations:
- CHC,
- clathrin heavy chain;
- tet,
- tetracycline
- Copyright © 2003, The National Academy of Sciences
