O-linked beta-N-acetylglucosaminyltransferase substrate specificity is regulated by myosin phosphatase targeting and other interacting proteins.
O-GlcNAc-transferase (OGT) substrate specificity is regulated by transiently interacting proteins. To further examine the regulation of OGT, we have identified 27 putative OGT-interacting proteins through a yeast two-hybrid screen. Two of these proteins, Trak1 (OIP106) and O-GlcNAcase, have been shown previously to interact with and regulate OGT. We demonstrate here that MYPT1 and CARM1 also interact with and target OGT. MYPT1 and CARM1 are substrates of OGT in vitro and in vivo. MYPT1 and CARM1 also function to alter OGT substrate specificity in vitro. Furthermore depletion of MYPT1 in Neuro-2a neuroblastoma cells alters GlcNAcylation of several proteins under basal conditions, suggesting that MYPT1 regulates OGT substrate specificity in vivo.
Cheung
Win D
WD
Department of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185, USA.
Sakabe
Kaoru
K
Housley
Michael P
MP
Dias
Wagner B
WB
Hart
Gerald W
GW
D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D002352:Carrier Proteins / Q000502:physiology*; D045744:Cell Line, Tumor; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D051379:Mice; D008954:Models, Biological; D009219:Myosin-Light-Chain Kinase / Q000502:physiology*; D043263:Myosin-Light-Chain Phosphatase / Q000737:chemistry*; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012319:RNA Polymerase II / Q000737:chemistry; D013379:Substrate Specificity; D020798:Two-Hybrid System Techniques
D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; C509169:Trak1 protein, mouse; D017351:N-Acetylglucosaminyltransferases; D009219:Myosin-Light-Chain Kinase; D012319:RNA Polymerase II; D043263:Myosin-Light-Chain Phosphatase; C510645:Ppp1r12a protein, mouse