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© The Rockefeller University Press, 0021-9525/1998//1559 $5.00
The Journal of Cell Biology, Volume 143, Number 6, , 1998 1559-1573

Role of Xklp3, a Subunit of the Xenopus Kinesin II Heterotrimeric Complex, in Membrane Transport between the Endoplasmic Reticulum and the Golgi Apparatus

Nathalie Le Bot^*, Claude Antony, Jamie White^*, Eric Karsenti^*, and Isabelle Vernos^*

^* Cell Biology and Biophysics Program, European Molecular Biological Laboratory, D-69117 Heidelberg, Germany; and Institut Curie, 75248 Paris Cedex 05, France

The function of the Golgi apparatus is to modify proteins and lipids synthesized in the ER and sort them to their final destination. The steady-state size and function of the Golgi apparatus is maintained through the recycling of some components back to the ER. Several lines of evidence indicate that the spatial segregation between the ER and the Golgi apparatus as well as trafficking between these two compartments require both microtubules and motors. We have cloned and characterized a new Xenopus kinesin like protein, Xklp3, a subunit of the heterotrimeric Kinesin II. By immunofluorescence it is found in the Golgi region. A more detailed analysis by EM shows that it is associated with a subset of membranes that contain the KDEL receptor and are localized between the ER and Golgi apparatus. An association of Xklp3 with the recycling compartment is further supported by a biochemical analysis and the behavior of Xklp3 in BFA-treated cells. The function of Xklp3 was analyzed by transfecting cells with a dominant-negative form lacking the motor domain. In these cells, the normal delivery of newly synthesized proteins to the Golgi apparatus is blocked. Taken together, these results indicate that Xklp3 is involved in the transport of tubular-vesicular elements between the ER and the Golgi apparatus.

Key Words: Kinesin II • microtubules • ER • Golgi • IC

Abbreviations used in this paper: AMP-PNP, 5'-adenylyllimido-diphosphate; BFA, brefeldin A; GFP, green fluorescent protein; GST, glutathione-S-transferase; HA, hemagglutinin; HP, Helix pomatia; IC, intermediate compartment; IFT, intraflagellar transport; KLP, kinesin-like protein; MBP, maltose-binding protein; MT, microtubules; PDI, protein disulfide isomerase; PNS, postnuclear supernatant.

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