TurboID functions as an efficient biotin ligase for BioID applications in Xenopus embryos

Kanzler, Charlotte R.; Donohue, Michael C.; Dowdle, Megan E.; Sheets, Michael

doi:10.1016/j.ydbio.2022.10.005

Cited by 6 publications

(4 citation statements)

References 28 publications

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“…In the studies above, data were normalized for TURBO‐ID expression, a logical approach that assumes that TURBO‐ID expression is dominant for total proteome labeling. This conclusion has been drawn in several different model organisms, including Xenopus laevis [31] . However, how the activity of TURBO‐ID varies depending on pH, ionic strength, and other microenvironment‐specific factors that could change across different cell types, is less well known.…”

Section: “Boo Was Our Neighbor He Gave Us Two Soap Dolls a Broken Wat...mentioning

confidence: 84%

“…This conclusion has been drawn in several different model organisms, including Xenopus laevis. [31] However, how the activity of TURBO-ID varies depending on pH, ionic strength, and other microenvironment-specific factors that could change across different cell types, is less well known. These are important parameters to define and will help better normalization protocols in the future.…”

Section: Crawling Into the Subject's Skin: Biotinylation-based Labeli...mentioning

confidence: 99%

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Climbing into their Skin to Understand Contextual Protein–Protein Associations and Localizations: Functional Investigations in Transgenic Live Model Organisms

Long,

Aye

2024

ChemBioChem

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Borrowing some quotes from Harper Lee's novel “To Kill A Mockingbird” to help frame our manuscript, we discuss methods to profile local proteomes. We initially focus on chemical biology regimens that function in live organisms and use reactive biotin species for this purpose. We then consider ways to add new dimensions to these experimental regimens, principally by releasing less reactive (i. e., more selective) (preter)natural electrophiles. Although electrophile release methods may have lower resolution and label fewer proteins than biotinylation methods, their ability to probe simultaneously protein function and locale raises new and interesting possibilities for the field.

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Section: “Boo Was Our Neighbor He Gave Us Two Soap Dolls a Broken Wat...mentioning

confidence: 84%

Section: Crawling Into the Subject's Skin: Biotinylation-based Labeli...mentioning

confidence: 99%

Climbing into their Skin to Understand Contextual Protein–Protein Associations and Localizations: Functional Investigations in Transgenic Live Model Organisms

Long,

Aye

2024

ChemBioChem

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“…Additionally, HRP is limited in its ability to carry out its labeling function depending upon cellular location, limiting its use for intracellular environments [ 25 ]. The benefits of using this updated BioID system allows for labeling of transient and weak interactions in minutes rather than hours without affecting cell viability, providing a record of interactions as the ligase moves through a cell [ 26 , 27 , 28 ]. Using the proximity labeling capabilities of TurboID, a TurboID-ER fusion construct was generated to profile ER interactomes.…”

Section: Introductionmentioning

confidence: 99%

TRIM33 Is a Co-Regulator of Estrogen Receptor Alpha

Romo,

Karakyriakou,

Cressey

et al. 2024

Cancers

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Estrogen receptor alpha (ER)-positive breast cancer is responsible for over 60% of breast cancer cases in the U.S. Among patients diagnosed with early-stage ER+ disease, 1/3 will experience recurrence despite treatment with adjuvant endocrine therapy. ER is a nuclear hormone receptor responsible for estrogen-driven tumor growth. ER transcriptional activity is modulated by interactions with coregulators. Dysregulation of the levels of these coregulators is involved in the development of endocrine resistance. To identify ER interactors that modulate transcriptional activity in breast cancer, we utilized biotin ligase proximity profiling of ER interactomes. Mass spectrometry analysis revealed tripartite motif containing 33 (TRIM33) as an estrogen-dependent interactor of ER. shRNA knockdown showed that TRIM33 promoted ER transcriptional activity and estrogen-induced cell growth. Despite its known role as an E3 ubiquitin ligase, TRIM33 increased the stability of endogenous ER in breast cancer cells. TRIM33 offers a novel target for inhibiting estrogen-induced cancer cell growth, particularly in cases of endocrine resistance driven by ER (ESR1) gene amplification or overexpression.

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“…Its labeling time can be shortened to 30 minutes, and it maintains catalytic activity at lower temperatures. Additionally, TurboID can label proteins in live physiological environments without disrupting the cells [21] . Here, we constructed PNO1-TurboID and NOB1-TurboID systems to use TurboID technology to identify the interaction proteome of PNO1 and NOB1 within cells.…”

Section: Introductionmentioning

confidence: 99%

the Interaction Proteome of Ribosomal 40S Components PNO1 and NOB1 Using TurboID Proximity Labeling Technology

Xu,

Zheng,

Chen

et al. 2024

Preprint

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Background The ribosome assembly factors PNO1 and NOB1 play crucial roles in the maturation of the 40S ribosomal small subunit. TurboID is an efficient biotin ligase that can biotinylate proteins in proximity to the target protein and is widely used to study complex biological processes within cells.Here, we utilized this technology to investigate the complex interaction network of PNO1 and NOB1 within cells. Results Through immunofluorescence experiments, we found that PNO1 and NOB1 have different localizations within cells. By identifying the proximal proteins biotinylated by PNO1-TurboID and NOB1-TurboID, we discovered 871 proteins interacting with PNO1 and 1044 proteins interacting with NOB1, with 663 proteins overlapping. These results suggest that PNO1 and NOB1 are extensively involved in various biological processes within the cell. Furthermore, we found that PNO1 and NOB1 interact with translation-related proteins EIF4B and EIF4G2, indicating that they may participate in the mRNA translation process. Conclusion The interaction proteome results of PNO1 and NOB1 suggest that ribosome assembly factors are not only involved in ribosome biogenesis but also couple with multiple biological processes within the cell, such as mRNA translation. This provides a foundation for understanding the complex biological processes within the cell.

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TurboID functions as an efficient biotin ligase for BioID applications in Xenopus embryos

Cited by 6 publications

References 28 publications

Climbing into their Skin to Understand Contextual Protein–Protein Associations and Localizations: Functional Investigations in Transgenic Live Model Organisms

Climbing into their Skin to Understand Contextual Protein–Protein Associations and Localizations: Functional Investigations in Transgenic Live Model Organisms

TRIM33 Is a Co-Regulator of Estrogen Receptor Alpha

the Interaction Proteome of Ribosomal 40S Components PNO1 and NOB1 Using TurboID Proximity Labeling Technology

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