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The O-β-linked N-acetylglucosaminylation of the Lamin B receptor and its impact on DNA binding and phosphorylation
- Source :
- Biochimica et Biophysica Acta (BBA)-General Subjects, Biochimica et Biophysica Acta (BBA)-General Subjects, Elsevier, 2018, 1862 (4), pp.825-835. ⟨10.1016/j.bbagen.2018.01.007⟩, Biochimica et Biophysica Acta (BBA)-General Subjects, 2018, 1862 (4), pp.825-835. ⟨10.1016/j.bbagen.2018.01.007⟩
- Publication Year :
- 2018
- Publisher :
- Elsevier BV, 2018.
-
Abstract
- Lamin B Receptor (LBR) is an integral protein of the interphase inner nuclear membrane that is implicated in chromatin anchorage to the nuclear envelope. Phosphorylation of a stretch of arginine-serine (RS) dipeptides in the amino-terminal nucleoplasmic domain of LBR regulates the interactions of the receptor with other nuclear proteins, DNA and RNA and thus modulates tethering of heterochromatin to the nuclear envelope. While phosphorylation has been extensively studied, very little is known about other post-translational modifications of the protein. There is only one report on the O-β-linked N-acetyl-glucosaminylation (O-GlcNAcylation) of a serine residue downstream of the RS domain of rat LBR. In the present study we identify additional O-GlcNAcylation sites by using as substrates of O-β-N-acetylglucosaminyltransferase (OGT) a set of peptides containing the entire LBR RS domain or parts of it as well as flanking sequences. The in vitro activity of OGT was assessed by tandem mass spectrometry and NMR spectroscopy. Furthermore, we provide evidence that O-GlcNAcylation hampers DNA binding while it marginally affects RS domain phosphorylation mediated by SRPK1, Akt2 and cdk1 kinases. General significance Our methodology providing a quantitative description of O-GlcNAc patterns based on a combination of mass spectrometry and high resolution NMR spectroscopy on short peptide substrates allows subsequent functional analyses. Hence, our approach is of general interest to a wide audience of biologists aiming at deciphering the functional role of O-GlcNAc glycosylation and its crosstalk with phosphorylation.
- Subjects :
- 0301 basic medicine
Turkeys
Glycosylation
Proto-Oncogene Proteins c-akt
[SDV]Life Sciences [q-bio]
Biophysics
Receptors, Cytoplasmic and Nuclear
Lamin B receptor
N-Acetylglucosaminyltransferases
Biochemistry
Acetylglucosamine
03 medical and health sciences
CDC2 Protein Kinase
Animals
Humans
Inner membrane
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Protein phosphorylation
Amino Acid Sequence
Phosphorylation
Nuclear protein
Molecular Biology
Integral membrane protein
ComputingMilieux_MISCELLANEOUS
Binding Sites
Sequence Homology, Amino Acid
Chemistry
DNA
Rats
Chromatin
Cell biology
030104 developmental biology
Peptides
Protein Binding
Subjects
Details
- ISSN :
- 03044165
- Volume :
- 1862
- Database :
- OpenAIRE
- Journal :
- Biochimica et Biophysica Acta (BBA) - General Subjects
- Accession number :
- edsair.doi.dedup.....590944fbeb0076397a171a6690212e73