Title : Dynamic interplay between O-glycosylation and O-phosphorylation of nucleocytoplasmic
proteins : alternative glycosylation/phosphorylation of
THR-58 , a known mutational hot spot of
c-Myc in lymphomas, is regulated by mitogens
Abstract :
- Previously, we reported that c-Myc is glycosylated by O-linked N-acetylglucosamine at Thr-58 , a known phosphorylation site and a mutational hot spot in lymphomas
- In this paper, we describe the production and characterization of two Thr-58 site-specific antibodies and use them to examine the modification of Thr-58 in living cells
- One antibody specifically reacts with the Thr-58-glycosylated form of c-Myc , and the other reacts only with unmodified Thr-58 in c-Myc
- Using these antibodies together with a commercial anti- Thr-58-phosphorylated c-Myc antibody, we simultaneously detected three forms of c-Myc ( Thr-58-unmodified, -phosphorylated, and -glycosylated)
- It has been reported that Thr-58 phosphorylation is dependent on a prior phosphorylation of Ser-62
- Mutagenesis of Ser-62 to Ala showed a marked decrease of Thr-58 phosphorylation and a marked increase of Thr-58 glycosylation
- Growth inhibition of HL60 cells by serum starvation increases Thr-58 glycosylation and correspondingly decreases its phosphorylation
- Serum stimulation has the opposite effect upon the modification status of Thr-58
- A candidate kinase responsible for Thr-58 phosphorylation is the glycogen synthase kinase 3 3 (GSK3)
- Lithium, a competitive inhibitor of GSK3, decreased Thr-58 phosphorylation and increased its glycosylation
- Finally, we show that the Thr-58-phosphorylated form of c-Myc predominantly accumulates in the cytoplasm rather than the nucleus upon inhibition of proteasome activity
- These data suggest that hierarchical phosphorylation of Ser-62 and Thr-58 and alternative glycosylation/phosphorylation of Thr-58 together regulate the myriad functions of c-Myc in cells
Output (sent_index, trigger,
protein,
sugar,
site):
- 0. O-glycosylation, , proteins, -, -
- 1. Thr-58, , -, O-linked N-acetylglucosamine, Thr-58
- 1. glycosylated, , c-Myc, -, -
- 3. Thr-58-glycosylated, , form of c-Myc, -, -
- 4. -glycosylated, , c-Myc, -, -
Output(Part-Of) (sent_index,
protein,
site):
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):
- 1. c-Myc, O-linked N-acetylglucosamine, Thr-58