title: Characterization of coding synonymous and non-synonymous variants in ADAMTS13 using ex vivo and in silico approaches.
abstract: Synonymous variations, which are defined as codon substitutions that do not change the encoded amino acid, were previously thought to have no effect on the properties of the synthesized protein(s). However, mounting evidence shows that these "silent" variations can have a significant impact on protein expression and function and should no longer be considered "silent". Here, the effects of six synonymous and six non-synonymous variations, previously found in the gene of ADAMTS13, the von Willebrand Factor (VWF) cleaving hemostatic protease, have been investigated using a variety of approaches. The ADAMTS13 mRNA and protein expression levels, as well as the conformation and activity of the variants have been compared to that of wild-type ADAMTS13. Interestingly, not only the non-synonymous variants but also the synonymous variants have been found to change the protein expression levels, conformation and function. Bioinformatic analysis of ADAMTS13 mRNA structure, amino acid conservation and codon usage allowed us to establish correlations between mRNA stability, RSCU, and intracellular protein expression. This study demonstrates that variants and more specifically, synonymous variants can have a substantial and definite effect on ADAMTS13 function and that bioinformatic analysis may allow development of predictive tools to identify variants that will have significant effects on the encoded protein.
authors: Laboratory of Hemostasis, Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.
authors: Nathan C
authors: NC
authors: Edwards
authors: Zachary A
authors: ZA
authors: Hing
authors: Avital
authors: A
authors: Perry
authors: Adam
authors: A
authors: Blaisdell
authors: David B
authors: DB
authors: Kopelman
authors: Robert
authors: R
authors: Fathke
authors: William
authors: W
authors: Plum
authors: Jordan
authors: J
authors: Newell
authors: Courtni E
authors: CE
authors: Allen
authors: Geetha
authors: G
authors: S
authors: Aaron
authors: A
authors: Shapiro
authors: Chinyere
authors: C
authors: Okunji
authors: Idit
authors: I
authors: Kosti
authors: Noam
authors: N
authors: Shomron
authors: Vahan
authors: V
authors: Grigoryan
authors: Teresa M
authors: TM
authors: Przytycka
authors: Zuben E
authors: ZE
authors: Sauna
authors: Raheleh
authors: R
authors: Salari
authors: Yael
authors: Y
authors: Mandel-Gutfreund
authors: Anton A
authors: AA
authors: Komar
authors: Chava
authors: C
authors: Kimchi-Sarfaty
mesh_terms: D051722:ADAM Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000071120:ADAMTS13 Protein; D019943:Amino Acid Substitution / Q000235:genetics*; D003062:Codon / Q000235:genetics*; D019295:Computational Biology / Q000379:methods*; D017124:Conserved Sequence / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D057809:HEK293 Cells; D006801:Humans; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017433:Protein Structure, Secondary; D059748:Proteolysis; D020871:RNA Stability / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity; D014357:Trypsin / Q000378:metabolism
chemical_list: D003062:Codon; D050505:Mutant Proteins; D012333:RNA, Messenger; D014357:Trypsin; D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human