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Vol. 11, Issue 1, 171-182, January 2000

Organization of the Yeast Golgi Complex into at Least Four Funtionally Distinct Compartments

William T. Brigance,^* Charles Barlowe, and Todd R. Graham^*

 ^*Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235; and  Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755-3844

Pro--factor (pro-f) is posttranslationally modified in the yeast Golgi complex by the addition of 1,6-, 1,2-, and 1,3-linked mannose to N-linked oligosaccharides and by a Kex2p-initiated proteolytic processing event. Previous work has indicated that the 1,6- and 1,3-mannosylation and Kex2p-dependent processing of pro-f are initiated in three distinct compartments of the Golgi complex. Here, we present evidence that 1,2-mannosylation of pro-f is also initiated in a distinct Golgi compartment. Linkage-specific antisera and an endo-1,6-D-mannanase (endoM) were used to quantitate the amount of each pro-f intermediate during transport through the Golgi complex. We found that 1,6-, 1,2-, and 1,3-mannose were sequentially added to pro-f in a temporally ordered manner, and that the intercompartmental transport factor Sec18p/N-ethylmaleimide-sensitive factor was required for each step. The Sec18p dependence implies that a transport event was required between each modification event. In addition, most of the Golgi-modified pro-f that accumulated in brefeldin A-treated cells received only 1,6-mannosylation as did ~50% of pro-f transported to the Golgi in vitro. This further supports the presence of an early Golgi compartment that houses an 1,6-mannosyltransferase but lacks 1,2-mannosyltransferase activity in vivo. We propose that the 1,6-, 1,2-, and 1,3-mannosylation and Kex2p-dependent processing events mark the cis, medial, trans, and trans-Golgi network of the yeast Golgi complex, respectively.

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Corresponding author. E-mail address: tr.graham{at}vanderbilt.edu.

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Molecular Biology of the Cell
Vol. 11, 171-182, January 2000
Copyright © 2000 by The American Society for Cell Biology

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