Caspase-mediated cleavage of the stacking protein GRASP65 is required for Golgi fragmentation during apoptosis

doi:10.1083/jcb.200110007

Published online 28 January 2002. doi:10.1083/jcb.200110007

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© The Rockefeller University Press, 0021-9525/2002/2/495 $5.00
The Journal of Cell Biology, Volume 156, Number 3, February 4, 2002 495-509

Article

Caspase-mediated cleavage of the stacking protein GRASP65 is required for Golgi fragmentation during apoptosis

Jon D. Lane¹, John Lucocq², James Pryde³, Francis A. Barr⁴, Philip G. Woodman¹, Victoria J. Allan¹ and Martin Lowe¹

¹ School of Biological Sciences, University of Manchester, Manchester M13 9PT, United Kingdom
² School of Life Sciences, WTB/MSI Complex, University of Dundee, Dundee DD1 5EH, United Kingdom
³ Department of Medical and Radiological Sciences, The University of Edinburgh, EH8 9YL Edinburgh, United Kingdom
⁴ Max-Planck Institute of Biochemistry, Department of Cell Biology, D-82152 Martinsried, Germany

Address correspondence to Martin Lowe, School of Biological Sciences, University of Manchester, 2.205 Stopford Bldg., Oxford Rd., Manchester M13 9PT, U.K. Tel.: 44-161-275-5387. Fax: 44-161-275-5082. E-mail: lowe{at}man.ac.uk

The mammalian Golgi complex is comprised of a ribbon of stackedcisternal membranes often located in the pericentriolar regionof the cell. Here, we report that during apoptosis the Golgiribbon is fragmented into dispersed clusters of tubulo-vesicularmembranes. We have found that fragmentation is caspase dependentand identified GRASP65 (Golgi reassembly and stacking proteinof 65 kD) as a novel caspase substrate. GRASP65 is cleaved specificallyby caspase-3 at conserved sites in its membrane distal COOHterminus at an early stage of the execution phase. Expressionof a caspase-resistant form of GRASP65 partially preserved cisternalstacking and inhibited breakdown of the Golgi ribbon in apoptoticcells. Our results suggest that GRASP65 is an important structuralcomponent required for maintenance of Golgi apparatus integrity.

Key Words: Golgi apparatus; apoptosis; GRASP65; Golgi structure; caspase

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