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Organelle specific O-glycosylation drives MMP14 activation, tumor growth, and metastasis

Cancer Cell. 2017; 
Anh Tuan Nguyen, Joanne Chia, Manon Ros, Kam Man Hui, Frederic Saltel, Frederic Bard
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Gene Synthesis To construct the sleeping beauty vector, the vector pT2/shp53/GFP4 was digested with XhoI and ligated to a 2176-bp XhoI/SalIsynthesized fragment by Genscript USA Inc.Galnt1 (or Golgi-G1), ER-G1 (fused to an ER signal sequence from human growth hormone), ER-G1 with catalytic domain mutations D156Q, D209N and H211D to block substrate and manganese binding were synthesized by GenScript USA Inc.To generate pT2/PGK/ mCherry-NRas-2A-MMP14-WT and pT2/PGK/mCherry-NRas-2A-MMP14-T(5)A vectors, human MMP14 wild-type and mutants containing 2A self-cleaving sequences were gene synthesized (Genscript), then cloned into vector pT2/PGK/mCherry-NRas by two SacII sites. The following vectors have been deposited at Addgene.To construct the pLENTI6.3 vectors, human GALNT1 (NM_020474) and human MMP14 (NM_004995) wild-type and mutants were gene synthesized (Genscript) and cloned into pDONR221 entry vector (Thermofisher scientific). T Get A Quote

摘要

Nguyen et al. find that O-glycosylation increases during liver tumor progression, with increased expression of GALNT1 and glycosylation of ER-associated proteins. In mouse models, expression of GALNT1 in the ER, but not the Golgi, accelerates tumorigenesis and increases invasion through glycosylation of MMP14.

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