Differential distribution of dynamin isoforms in mammalian cells

Hong Cao, Fabiana Garcia, Mark A Mc Niven

Research output: Contribution to journalArticle

310 Citations (Scopus)

Abstract

Dynamins are 100-kDa GTPases that are essential for clathrin-coated vesicle formation during receptor-mediated endocytosis. To date, three different dynamin genes have been identified, with each gene expressing at least four different alternatively spliced forms. Currently, it is unclear whether these different dynamin gene products perform distinct or redundant cellular functions. Therefore, the focus of this study was to identify additional spliced variants of dynamin from rat tissues and to define the distribution of the dynamin family members in a cultured rat epithelial cell model (Clone 9 cells). After long-distance reverse transcription (RT)-PCR of mRNA from different rat tissues, the full-length cDNAs encoding the different dynamin isoforms were sequenced and revealed four additional spliced variants for dynamin I and nine for dynamin III. Thus, in rat tissues there are a total of at least 25 different mRNAs produced from the three dynamin genes. Subsequently, we generated stably transfected Clone 9 cells expressing full- length cDNAs of six different spliced forms tagged with green fluorescent protein. Confocal or fluorescence microscopy of these transfected cells revealed that many of the dynamin proteins associate with distinct membrane compartments, which include clathrin-coated pits at the plasma membrane and the Golgi apparatus, and several undefined vesicle populations. These results indicate that the dynamin family is more extensive than was originally predicted and suggest that the different dynamin proteins are localized to distinct cytoplasmic or membrane compartments.

Original languageEnglish (US)
Pages (from-to)2595-2609
Number of pages15
JournalMolecular Biology of the Cell
Volume9
Issue number9
StatePublished - Sep 1998

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Dynamins
Protein Isoforms
Dynamin III
Genes
Dynamin I
Complementary DNA
Clone Cells
Cell Membrane
Clathrin-Coated Vesicles
Clathrin
Messenger RNA
GTP Phosphohydrolases
Golgi Apparatus
Endocytosis
Green Fluorescent Proteins
Fluorescence Microscopy
Confocal Microscopy
Reverse Transcription
Proteins
Epithelial Cells

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Differential distribution of dynamin isoforms in mammalian cells. / Cao, Hong; Garcia, Fabiana; Mc Niven, Mark A.

In: Molecular Biology of the Cell, Vol. 9, No. 9, 09.1998, p. 2595-2609.

Research output: Contribution to journalArticle

Cao, Hong ; Garcia, Fabiana ; Mc Niven, Mark A. / Differential distribution of dynamin isoforms in mammalian cells. In: Molecular Biology of the Cell. 1998 ; Vol. 9, No. 9. pp. 2595-2609.
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