Mitochondrial and nucleocytoplasmic targeting of O-linked GlcNAc transferase

Journal of Cell Science - Tập 116 Số 4 - Trang 647-654 - 2003
Dona C. Love1, Jarema Kochran2,3, R. Lamont Cathey1,4, Sanghoon Shin1, John A. Hanover1
1Laboratory of Cell Biochemistry and Biology, NIDDK, National Institutes of Health, Bethesda, MD, 20892, USA
2Department of Metabolic Diseases, Hoffman-La Roche Inc., 340 Kingsland Street,Nutley, NJ, 07110, USA
3NATIONAL INSTITUTES OF HEALTH
4Present address: Department of General Surgery, Carolinas Medical Center,Charlotte, NC 28232-2861, USA

Tóm tắt

O-linked GlcNAc transferase (OGT) mediates a novel glycan-dependent signaling pathway, but the intracellular targeting of OGT is poorly understood. We examined the localization of OGT by immunofluorescence microscopy, subcellular fractionation and immunoblotting using highly specific affinity-purified antisera. In addition to the expected nuclear localization,we found that OGT was highly concentrated in mitochondria. Since the mitochondrial OGT (103 kDa) was smaller than OGT found in other compartments(116 kDa) we reasoned that it was one of two predicted splice variants of OGT. The N-termini of these isoforms are unique; the shorter form contains a potential mitochondrial targeting sequence. We found that when epitope-tagged,the shorter form (mOGT; 103 kDa) concentrated in HeLa cell mitochondria,whereas the longer form (ncOGT; 116 kDa) localized to the nucleus and cytoplasm. The N-terminus of mOGT was essential for proper targeting. Although mOGT appears to be an active transferase, O-linked GlcNAc-modified substrates do not accumulate in mitochondria. Using immunoelectron microscopy and mitochondrial fractionation, we found that mOGT was tightly associated with the mitochondrial inner membrane. The differential localization of mitochondrial and nucleocytoplasmic isoforms of OGT suggests that they perform unique intracellular functions.

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Journal of Cell Science

Tập 116 Số 4

647-654

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