title: The melanoma-associated antigen 1 (MAGEA1) protein stimulates the E3 ubiquitin-ligase activity of TRIM31 within a TRIM31-MAGEA1-NSE4 complex.
abstract: The MAGE (Melanoma-associated antigen) protein family members are structurally related to each other by a MAGE-homology domain comprised of 2 winged helix motifs WH/A and WH/B. This family specifically evolved in placental mammals although single homologs designated NSE3 (non-SMC element) exist in most eukaryotes. NSE3, together with its partner proteins NSE1 and NSE4 form a tight subcomplex of the structural maintenance of chromosomes SMC5-6 complex. Previously, we showed that interactions of the WH/B motif of the MAGE proteins with their NSE4/EID partners are evolutionarily conserved (including the MAGEA1-NSE4 interaction). In contrast, the interaction of the WH/A motif of NSE3 with NSE1 diverged in the MAGE paralogs. We hypothesized that the MAGE paralogs acquired new RING-finger-containing partners through their evolution and form MAGE complexes reminiscent of NSE1-NSE3-NSE4 trimers. In this work, we employed the yeast 2-hybrid system to screen a human RING-finger protein library against several MAGE baits. We identified a number of potential MAGE-RING interactions and confirmed several of them (MDM4, PCGF6, RNF166, TRAF6, TRIM8, TRIM31, TRIM41) in co-immunoprecipitation experiments. Among these MAGE-RING pairs, we chose to examine MAGEA1-TRIM31 in detail and showed that both WH/A and WH/B motifs of MAGEA1 bind to the coiled-coil domain of TRIM31 and that MAGEA1 interaction stimulates TRIM31 ubiquitin-ligase activity. In addition, TRIM31 directly binds to NSE4, suggesting the existence of a TRIM31-MAGEA1-NSE4 complex reminiscent of the NSE1-NSE3-NSE4 trimer. These results suggest that MAGEA1 functions as a co-factor of TRIM31 ubiquitin-ligase and that the TRIM31-MAGEA1-NSE4 complex may have evolved from an ancestral NSE1-NSE3-NSE4 complex.
authors.affiliation: a From the Mendel Center for Plant Genomics and Proteomics; Central European Institute of Technology; Masaryk University ; Brno , Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Kozakova
authors.forename: Lucie
authors.initials: L
authors.lastname: Vondrova
authors.forename: Karel
authors.initials: K
authors.lastname: Stejskal
authors.forename: Panagoula
authors.initials: P
authors.lastname: Charalabous
authors.forename: Peter
authors.initials: P
authors.lastname: Kolesar
authors.forename: Alan R
authors.initials: AR
authors.lastname: Lehmann
authors.forename: Stjepan
authors.initials: S
authors.lastname: Uldrijan
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Sanderson
authors.forename: Zbynek
authors.initials: Z
authors.lastname: Zdrahal
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Palecek
mesh_terms: D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D002853:Chromatography, Liquid; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D008954:Models, Biological; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D055503:Protein Multimerization; D054829:RING Finger Domains; D053719:Tandem Mass Spectrometry; D000070584:Tripartite Motif Proteins; D020798:Two-Hybrid System Techniques; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; C506530:MAGE-A1 protein (278-286), human; D046912:Multiprotein Complexes; C000603520:NSE4 protein, human; C533270:NSMCE1 protein, human; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D010455:Peptides; D000070584:Tripartite Motif Proteins; C534233:TRIM31 protein, human; D044767:Ubiquitin-Protein Ligases
keywords: E3 ubiquitin ligase; MAGE, melanoma-associated antigen; MAGEA1; MDM4; MHD, MAGE homology domain; NSE, non-SMC element; NSE1-NSE3-NSE4 complex; NSE4/EID family; PCGF6; RING-finger proteins; RNF166; SMC, structure maintenance of chromosomes; TRAF6; TRIM family; TRIM, tripartite motif; TRIM31; TRIM41; TRIM8; WH, winged helix; Y2H, yeast 2-hybrid.; melanoma-associated antigen family; protein evolution; ubiquitination