Resonance assignment and secondary structure of the tandem harmonin homology domains of human RTEL1

Kumar, Niranjan; Ghosh, Meenakshi; Manikandan, Parthasarathy; Basak, Sanmoyee; Akula, Deepa; Singh, Mahavir

doi:10.1007/s12104-022-10074-7

Cited by 3 publications

(7 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The secondary structures match well with the NMR chemical shift indexing-based secondary structures (Kumar et al, 2022). The overall topology of RTEL1 HHD2 is similar to the Nterminal domain of the human Harmonin (Faure et al, 2014).…”

Section: Structural Basis Of Rtel1 Hhd2 and Rpa 32c Interactionsupporting

confidence: 63%

“…All the helices are mainly a-helical (3.613) except the N-and C-termini of H4 and the N-terminal of H5, which consists of short 310 helices (Figure 2C and 2D). The secondary structures match well with the NMR chemical shift indexing-based secondary structures (Kumar et al, 2022). The overall topology of RTEL1 HHD2 is similar to the N-terminal domain of the human Harmonin (Faure et al, 2014).…”

Section: Structural Basis Of Rtel1 Hhd2 and Rpa 32c Interactionsupporting

confidence: 63%

“…RTEL1 HHD1 and HHD2 domains were expressed and purified as described previously (Kumar et al, 2022). In summary, the following protocol was followed for the purification of HHD1, HHD2, and HHD1+2.…”

Section: Methodsmentioning

confidence: 99%

“…Our recently deposited chemical shifts of HHD2 (BMRB entry 51077) (Kumar et al, 2022) were used to map the RPA 32C binding surface on the HHD2 crystal structure. The observed CSPs in the titration of 15 N-labelled HHD2 with 32C (Figure 2A) were quantified and plotted (Figure 3A).…”

Section: Structural Basis Of Rtel1 Hhd2 and Rpa 32c Interactionmentioning

confidence: 99%

“…The electrostatic surface potential of HHD2 revealed a distinct pattern of positively and negatively charged surfaces (Figure 4D). Our recently deposited chemical shifts of HHD2 (BMRB entry 51077) (Kumar et al, 2022) were used to map the RPA 32C binding surface on the HHD2 crystal structure. The observed CSPs in the titration of 15 N-labelled HHD2 with 32C (Figure 3A) were calculated and plotted (Figure 5A).…”

Section: Structural Basis Of Rtel1 Hhd2 and Rpa 32c Interactionmentioning

confidence: 99%

See 4 more Smart Citations

Harmonin homology domain-mediated interaction of RTEL1 helicase with RPA and DNA provides mechanistic insight into its role in DNA repair

Kumar

Taneja

Ghosh

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Regulator of telomere elongation helicase 1 (RTEL1) is an Fe-S cluster containing helicase that plays important roles in telomere DNA maintenance, DNA repair, and genome stability. It is a modular protein comprising a helicase domain, two tandem harmonin homology domains 1 & 2 (HHD1 and HHD2), and a RING domain. The RING domain interacts with telomere DNA binding protein TRF2 and helps in the recruitment of RTEL1 to the telomeres while the helicase domain disassembles telomere DNA t/D-loop and unwinds G-quadruplexes. The novel HHDs are predicted to mediate protein-protein interactions. Mutations in RTEL1 are associated with Hoyeraal-Hreidarsson syndrome and familial pulmonary fibrosis. Several of these mutations are found in the HHDs of RTEL1. In this study, we have carried out the structural characterization of HHDs of RTEL1. We report that human RTEL1 interacts and co-localizes with the replication protein A (RPA) in the cell. Using NMR spectroscopy and ITC experiments, we have shown that the HHD2 of RTEL1 interacts directly with the winged-helix 32C domain of RPA as well as DNA using an overlapping surface. These results establish an interplay among RTEL1, RPA, and DNA that has implications for understanding the role of RTEL1 in DNA replication, repair, and recombination processes.

show abstract

Section: Structural Basis Of Rtel1 Hhd2 and Rpa 32c Interactionsupporting

confidence: 63%

Section: Structural Basis Of Rtel1 Hhd2 and Rpa 32c Interactionsupporting

confidence: 63%

Section: Methodsmentioning

confidence: 99%

Section: Structural Basis Of Rtel1 Hhd2 and Rpa 32c Interactionmentioning

confidence: 99%

Section: Structural Basis Of Rtel1 Hhd2 and Rpa 32c Interactionmentioning

confidence: 99%

See 3 more Smart Citations

Harmonin homology domain-mediated interaction of RTEL1 helicase with RPA and DNA provides mechanistic insight into its role in DNA repair

Kumar

Taneja

Ghosh

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Harmonin homology domain-mediated interaction of RTEL1 helicase with RPA and DNA provides insights into its recruitment to DNA repair sites

Kumar,

Taneja,

Ghosh

et al. 2023

Nucleic Acids Research

View full text Add to dashboard Cite

The regulator of telomere elongation helicase 1 (RTEL1) plays roles in telomere DNA maintenance, DNA repair, and genome stability by dismantling D-loops and unwinding G-quadruplex structures. RTEL1 comprises a helicase domain, two tandem harmonin homology domains 1&2 (HHD1 and HHD2), and a Zn2+-binding RING domain. In vitro D-loop disassembly by RTEL1 is enhanced in the presence of replication protein A (RPA). However, the mechanism of RTEL1 recruitment at non-telomeric D-loops remains unknown. In this study, we have unravelled a direct physical interaction between RTEL1 and RPA. Under DNA damage conditions, we showed that RTEL1 and RPA colocalise in the cell. Coimmunoprecipitation showed that RTEL1 and RPA interact, and the deletion of HHDs of RTEL1 significantly reduced this interaction. NMR chemical shift perturbations (CSPs) showed that RPA uses its 32C domain to interact with the HHD2 of RTEL1. Interestingly, HHD2 also interacted with DNA in the in vitro experiments. HHD2 structure was determined using X-ray crystallography, and NMR CSPs mapping revealed that both RPA 32C and DNA competitively bind to HHD2 on an overlapping surface. These results establish novel roles of accessory HHDs in RTEL1’s functions and provide mechanistic insights into the RPA-mediated recruitment of RTEL1 to DNA repair sites.

show abstract

Discovery of a Novel Shared Variant Among RTEL1 Gene and RTEL1-TNFRSF6B lncRNA at Chromosome 20q13.33 in Familial Progressive Myoclonus Epilepsy

Chaudhari,

Acharya,

Jasti

et al. 2024

International Journal of Genomics

View full text Add to dashboard Cite

Background: Progressive myoclonus epilepsy (PME) is a neurodegenerative disorder marked by recurrent seizures and progressive myoclonus. To date, based on the phenotypes and causal genes, more than 40 subtypes of PMEs have been identified, and more remain to be characterized. Our study is aimed at identifying the aberrant gene(s) possibly associated with PMEs in two siblings born to asymptomatic parents, in the absence of known genetic mutations.Methods: Clinical assessments and molecular analyses, such as the repeat expansion test for CSTB; SCA1, 2, 3, 6, and 7; whole exome sequencing (WES); and mitochondrial genome sequencing coupled with computational analysis, were performed.Results: A family‐based segregation analysis of WES data was performed to identify novel genes associated with PMEs. The potassium channel, KCNH8 [c.298T>C; (p.Tyr100His)], a DNA repair gene, regulator of telomere elongation helicase 1 (RTEL1) [c.691G>T; (p.Asp231Tyr)] and long noncoding RNA, RTEL1-TNFRSF6B [chr20:62298898_G>T; NR_037882.1, hg19] were among the candidate genes that were found to be associated with PMEs. These homozygous variations in siblings belong to genes with a loss‐of‐function intolerant (pLI) score of ≤ 0.86, expected to be detrimental by multiple computational analyses, and were heterozygous in parents. Additionally, computational analysis and the expression of RTEL1 and RTEL1-TNFRSF6B revealed that RTEL1-TNFRSF6B may modulate RTEL1 via hsa‐miR‐3529‐3p. In the patient with the severe phenotype, a further deleterious mutation in SLC22A17 was identified. No de novo variants specific to these probands were identified in the mitochondrial genome.Conclusions: Our study is the first to report variants in KCNH8, RTEL1, and RTEL1-TNFRSF6B among PME cases. These genes when characterized fully may shed light on pathogenicity and have the potential to be used in the diagnosis of PME.

show abstract

Resonance assignment and secondary structure of the tandem harmonin homology domains of human RTEL1

Cited by 3 publications

References 21 publications

Harmonin homology domain-mediated interaction of RTEL1 helicase with RPA and DNA provides mechanistic insight into its role in DNA repair

Harmonin homology domain-mediated interaction of RTEL1 helicase with RPA and DNA provides mechanistic insight into its role in DNA repair

Harmonin homology domain-mediated interaction of RTEL1 helicase with RPA and DNA provides insights into its recruitment to DNA repair sites

Discovery of a Novel Shared Variant Among RTEL1 Gene and RTEL1-TNFRSF6B lncRNA at Chromosome 20q13.33 in Familial Progressive Myoclonus Epilepsy

Contact Info

Product

Resources

About