Identification of Ku70 Domain-Specific Interactors Using BioID2
Since its inception, proximity-dependent biotin identification (BioID), an in vivo biochemical screening method to identify proximal protein interactors, has seen extensive developments. Improvements and variants of the original BioID technique are being reported regularly, each expanding upon the e...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-03-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/10/3/646 |
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| author | Sanna Abbasi Caroline Schild-Poulter |
| author_facet | Sanna Abbasi Caroline Schild-Poulter |
| author_sort | Sanna Abbasi |
| collection | DOAJ |
| description | Since its inception, proximity-dependent biotin identification (BioID), an in vivo biochemical screening method to identify proximal protein interactors, has seen extensive developments. Improvements and variants of the original BioID technique are being reported regularly, each expanding upon the existing potential of the original technique. While this is advancing our capabilities to study protein interactions under different contexts, we have yet to explore the full potential of the existing BioID variants already at our disposal. Here, we used BioID2 in an innovative manner to identify and map domain-specific protein interactions for the human Ku70 protein. Four HEK293 cell lines were created, each stably expressing various BioID2-tagged Ku70 segments designed to collectively identify factors that interact with different regions of Ku70. Historically, although many interactions have been mapped to the C-terminus of the Ku70 protein, few have been mapped to the N-terminal von Willebrand A-like domain, a canonical protein-binding domain ideally situated as a site for protein interaction. Using this segmented approach, we were able to identify domain-specific interactors as well as evaluate advantages and drawbacks of the BioID2 technique. Our study identifies several potential new Ku70 interactors and validates RNF113A and Spindly as proteins that contact or co-localize with Ku in a Ku70 vWA domain-specific manner. |
| first_indexed | 2024-03-10T13:15:10Z |
| format | Article |
| id | doaj.art-1b67abd26c55425284831a475b7c36b7 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T13:15:10Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-1b67abd26c55425284831a475b7c36b72023-11-21T10:28:07ZengMDPI AGCells2073-44092021-03-0110364610.3390/cells10030646Identification of Ku70 Domain-Specific Interactors Using BioID2Sanna Abbasi0Caroline Schild-Poulter1Robarts Research Institute and Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON N6A 5B7, CanadaRobarts Research Institute and Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON N6A 5B7, CanadaSince its inception, proximity-dependent biotin identification (BioID), an in vivo biochemical screening method to identify proximal protein interactors, has seen extensive developments. Improvements and variants of the original BioID technique are being reported regularly, each expanding upon the existing potential of the original technique. While this is advancing our capabilities to study protein interactions under different contexts, we have yet to explore the full potential of the existing BioID variants already at our disposal. Here, we used BioID2 in an innovative manner to identify and map domain-specific protein interactions for the human Ku70 protein. Four HEK293 cell lines were created, each stably expressing various BioID2-tagged Ku70 segments designed to collectively identify factors that interact with different regions of Ku70. Historically, although many interactions have been mapped to the C-terminus of the Ku70 protein, few have been mapped to the N-terminal von Willebrand A-like domain, a canonical protein-binding domain ideally situated as a site for protein interaction. Using this segmented approach, we were able to identify domain-specific interactors as well as evaluate advantages and drawbacks of the BioID2 technique. Our study identifies several potential new Ku70 interactors and validates RNF113A and Spindly as proteins that contact or co-localize with Ku in a Ku70 vWA domain-specific manner.https://www.mdpi.com/2073-4409/10/3/646BioID2proximity-dependent biotin identificationKu70von Willebrand A-like domainRNF113ASpindly |
| spellingShingle | Sanna Abbasi Caroline Schild-Poulter Identification of Ku70 Domain-Specific Interactors Using BioID2 Cells BioID2 proximity-dependent biotin identification Ku70 von Willebrand A-like domain RNF113A Spindly |
| title | Identification of Ku70 Domain-Specific Interactors Using BioID2 |
| title_full | Identification of Ku70 Domain-Specific Interactors Using BioID2 |
| title_fullStr | Identification of Ku70 Domain-Specific Interactors Using BioID2 |
| title_full_unstemmed | Identification of Ku70 Domain-Specific Interactors Using BioID2 |
| title_short | Identification of Ku70 Domain-Specific Interactors Using BioID2 |
| title_sort | identification of ku70 domain specific interactors using bioid2 |
| topic | BioID2 proximity-dependent biotin identification Ku70 von Willebrand A-like domain RNF113A Spindly |
| url | https://www.mdpi.com/2073-4409/10/3/646 |
| work_keys_str_mv | AT sannaabbasi identificationofku70domainspecificinteractorsusingbioid2 AT carolineschildpoulter identificationofku70domainspecificinteractorsusingbioid2 |