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Structural and biochemical characterization of the C‐terminal region of the human RTEL 1 helicase

RTEL1 is an essential DNA helicase which plays an important role in various aspects of genome stability, from telomere metabolism to DNA replication, repair and recombination. RTEL1 has been implicated in a number of genetic diseases and cancer development, including glioma, breast, lung and gastroi...

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Bibliographic Details
Published in:Protein science 2024-09, Vol.33 (9)
Main Authors: Cortone, Giuseppe, Graewert, Melissa A., Kanade, Manil, Longo, Antonio, Hegde, Raghurama, González‐Magaña, Amaia, Chaves‐Arquero, Belén, Blanco, Francisco J., Napolitano, Luisa M. R., Onesti, Silvia
Format: Article
Language:English
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Summary:RTEL1 is an essential DNA helicase which plays an important role in various aspects of genome stability, from telomere metabolism to DNA replication, repair and recombination. RTEL1 has been implicated in a number of genetic diseases and cancer development, including glioma, breast, lung and gastrointestinal tumors. RTEL1 is a FeS helicase but, in addition to the helicase core, it comprises a long C‐terminal region which includes a number of folded domains connected by intrinsically disordered loops and mediates RTEL1 interaction with factors involved in pivotal cellular pathways. However, information on the architecture and the function of this region is still limited. We expressed and purified a variety of fragments encompassing the folded domains and the unstructured regions. We determined the crystal structure of the second repeat, confirming that it has a fold similar to the harmonin homology domains. SAXS data provide low‐resolution information on all the fragments and suggest that the presence of the RING domain affects the overall architecture of the C‐terminal region, making the structure significantly more compact. NMR data provide experimental information on the interaction between PCNA and the RTEL1 C‐terminal region, revealing a putative low‐affinity additional site of interaction. A biochemical analysis shows that the C‐terminal region, in addition to a preference for telomeric RNA and DNA G‐quadruplexes, has a high affinity for R‐loops and D‐loops, consistent with the role played by the RTEL1 helicase in homologous recombination, telomere maintenance and preventing replication‐transcription conflicts. We further dissected the contribution of each domain in binding different substrates.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.5093