A G-quadruplex DNA structure resolvase, RHAU, is essential for spermatogonia differentiation

Gao, Xiang; Ma, Wei; Nie, Junwei; Zhang, C; Zhang, Jiahan; Yao, G.-X.; Han, Jinxiong; Xu, Juan; Hu, Boqiang; Du, Yongkun; Shi, Qinghua; Yang, Zhongzhou; Huang, Xinyi; Zhang, Y

doi:10.1038/cddis.2014.571

Cited by 31 publications

(37 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the previous study, we found that MS-275 stimulates bone formation by interaction with Dhx36, (13) also known for its critical roles in the process of cell proliferation. (38) Therefore, we deduced that Dhx36mediated signaling might be participating in highly triggered cell proliferative activity through MS-275 administration. Considering the significance of proliferation of osteoblast progenitor cells in the calvarial suture region during development, (29) it could be the critical distinction of MS-275 treatment toward CCD.…”

Section: Discussionmentioning

confidence: 98%

An HDAC Inhibitor, Entinostat/MS-275, Partially Prevents Delayed Cranial Suture Closure in Heterozygous Runx2 Null Mice

Bae

Yoon

Cho

et al. 2016

Journal of Bone and Mineral Research

View full text Add to dashboard Cite

Cleidocranial dysplasia (CCD) is an autosomal dominant skeletal disorder caused by mutations in RUNX2, coding a key transcription factor of early osteogenesis. CCD patients suffer from developmental defects in cranial bones. Despite numerous investigations and clinical approaches, no therapeutic strategy has been suggested to prevent CCD. Here, we show that fetal administration of Entinostat/MS-275, a class I histone deacetylase (HDAC)-specific inhibitor, partially prevents delayed closure of cranial sutures in Runx2 þ/-mice strain of C57BL/6J by two mechanisms: 1) posttranslational acetylation of Runx2 protein, which stabilized the protein and activated its transcriptional activity; and 2) epigenetic regulation of Runx2 and other bone marker genes. Moreover, we show that MS-275 stimulates osteoblast proliferation effectively both in vivo and in vitro, suggesting that delayed skeletal development in CCD is closely related to the decreased number of progenitor cells as well as the delayed osteogenic differentiation. These findings provide the potential benefits of the therapeutic strategy using MS-275 to prevent CCD.

show abstract

Section: Discussionmentioning

confidence: 98%

An HDAC Inhibitor, Entinostat/MS-275, Partially Prevents Delayed Cranial Suture Closure in Heterozygous Runx2 Null Mice

Bae

Yoon

Cho

et al. 2016

Journal of Bone and Mineral Research

View full text Add to dashboard Cite

show abstract

“…RHAU helicase, also named G4 resolvase 1 (DHX36 gene product, or G4R1, from human) is a superfamily 2 DEAH-box helicase that plays a role in heart development (11), hematopoiesis (12) and spermatogenesis (13). RHAU was originally identified as a RNA helicase that binds RNA G4 with sub-nanomolar affinity (14–17).…”

Section: Introductionmentioning

confidence: 99%

“…RHAU has also been shown to bind DNA G4 with similar affinity to RNA and can unfold DNA G4 (24), which also has crucial biological functions. In addition, a recent study revealed that RHAU directly binds to the DNA G4 structures formed on the promoter region of c-Kit , facilitating its expression and regulating spermatogonia differentiation (13). Sequence analysis showed that almost all metazoan species have RHAU orthologues (25).…”

Section: Introductionmentioning

confidence: 99%

RHAU helicase stabilizes G4 in its nucleotide-free state and destabilizes G4 upon ATP hydrolysis

et al. 2016

View full text Add to dashboard Cite

The DEAH-box ATP-dependent RHAU helicases specifically unfold RNA and DNA G-quadruplexes (G4s). However, it remains unclear how the RHAU's G4 unfolding activity is coupled to different stages of the ATPase cycle. Here, using a single-molecule manipulation approach, we show that binding of Drosophila RHAU stabilizes an intramolecularly folded parallel DNA G4 against mechanical unfolding in its nucleotide-free and in its AMP-PNP or ADP bound states, while it destabilizes the G4 when coupled to ATP hydrolysis. Importantly, our results show that the ADP·AlF\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} }{}$_4^-$\end{document}-bound RHAU does not stabilize the G4. We also found that both a single-stranded 3′ DNA tail and the RSM domain of RHAU that binds specifically to the G4 structure, are dispensable for the stabilization of the G4, but both are required for G4 destabilization. Our study provides the first evidence that the unfolding kinetics of a G-quadruplex can be modulated by different nucleotide-bound states of the helicase.

show abstract

“…DHX36 has been implicated as a regulator of RNA folding and assembly (17,18), RNA localization, translation (19), pre-mRNA processing (20), and transcription (21), most simply suggesting that it functions in both the cytoplasm and nucleus and interacts with both RNA and DNA G4s. Underscoring the importance of this protein and potentially G4s in vivo, DHX36 is an essential gene in mouse (22)(23)(24).…”

Section: ___________________________________mentioning

confidence: 99%

The G-quadruplex (G4) resolvase DHX36 efficiently and specifically disrupts DNA G4s via a translocation-based helicase mechanism

Yangyuoru

Bradburn

et al. 2018

Journal of Biological Chemistry

View full text Add to dashboard Cite

Single-stranded DNA (ssDNA) and RNA regions that include at least four closely spaced runs of three or more consecutive guanosines strongly tend to fold into stable G-quadruplexes (G4s). G4s play key roles as DNA regulatory sites and as kinetic traps that can inhibit biological processes, but how G4s are regulated in cells remains largely unknown. Here, we developed a kinetic framework for G4 disruption by DEAH-box helicase 36 (DHX36), the dominant G4 resolvase in human cells. Using tetramolecular DNA and RNA G4s with four to six G-quartets, we found that DHX36-mediated disruption is highly efficient, with rates that depend on G4 length under saturating conditions (k cat ) but not under subsaturating conditions (k cat /K M ). These results suggest that a step during G4 disruption limits the k cat value and that DHX36 binding limits k cat /K M . Similar results were obtained for unimolecular DNA G4s. DHX36 activity depended on a 3′ ssDNA extension and was blocked by a polyethylene glycol linker, indicating that DHX36 loads onto the extension and translocates 3′-5′ toward the G4. DHX36 unwound dsDNA poorly compared with G4s of comparable intrinsic lifetime. Interestingly, we observed that DHX36 has striking 3′-extension sequence preferences that differ for G4 disruption and dsDNA unwinding, most likely arising from differences in the rate-limiting step for the two activities. Our results indicate that DHX36 disrupts G4s with a conventional helicase mechanism that is tuned for great efficiency and specificity for G4s. The dependence of DHX36 on the 3′-extension sequence suggests that the extent of formation of genomic G4s may not track directly with G4 stability. ___________________________________Single-stranded DNA and RNA segments that include at least four closely-spaced runs of three or more consecutive guanosine nucleotides have a strong intrinsic propensity to fold into stable G-quadruplex structures (G4s) (1) (Fig. 1A, top left). The genomes of humans and many other eukaryotes are replete with putative G4-forming sequences, and pronounced, conserved patterns have been observed in their distribution, suggesting that Gquadruplex structures form at least transiently in cells and perform conserved functions (2-6). Supporting this hypothesis, G4s have been detected in cells for both .To function as regulatory elements, G4s must be folded and disrupted in a regulated way. In addition, processes that require single-stranded DNA or RNA, such as replication and translation, require that G4s be temporarily disrupted. The high stability and long intrinsic lifetime of G4s suggest Mechanism of G4 disruption by DHX362 that their efficient disruption requires the activity of ATP-dependent enzymes such as helicases.Supporting this view, incorporation of sequences predicted to form particularly stable G4s leads to genetic instability in yeast, suggesting that these G4 structures are not processed efficiently and pose blocks to replication (12). Indeed, several helicases have been shown to possess G4 disruption acti...

show abstract

A G-quadruplex DNA structure resolvase, RHAU, is essential for spermatogonia differentiation

Cited by 31 publications

References 68 publications

An HDAC Inhibitor, Entinostat/MS-275, Partially Prevents Delayed Cranial Suture Closure in Heterozygous Runx2 Null Mice

An HDAC Inhibitor, Entinostat/MS-275, Partially Prevents Delayed Cranial Suture Closure in Heterozygous Runx2 Null Mice

RHAU helicase stabilizes G4 in its nucleotide-free state and destabilizes G4 upon ATP hydrolysis

The G-quadruplex (G4) resolvase DHX36 efficiently and specifically disrupts DNA G4s via a translocation-based helicase mechanism

Contact Info

Product

Resources

About