Functional genomic screen for modulators of ciliogenesis and cilium length

Kim, Joon; Lee, Ji Eun; Heynen‐Genel, Susanne; Suyama, Eigo; Ono, Keiichiro; Lee, KiYoung; Ideker, Trey; Aza-Blanc, Pedro; Gleeson, Joseph G.

doi:10.1038/nature08895

Cited by 487 publications

(674 citation statements)

References 24 publications

Supporting

Mentioning

612

Contrasting

Unclassified

Order By: Relevance

“…It is likely that a variety of pathways contribute to modifications of cilia length and structure. 16,22 Cyclic loading of the rat tail tendons in the current study was able to maintain cilia length (with respect to fresh controls) in tendons loaded immediately following harvest and reverse (shorten) cilia lengthened in response to SD, returning them to control lengths. Previous studies from our lab have shown that the loading scheme utilized in the current study (3% strain at 0.17 Hz) were able to maintain tendon cell homeostasis with respect to catabolic gene expression 27 or decrease catabolic gene expression in 24 h SD tendons.…”

Section: Discussionmentioning

confidence: 96%

“…Cilia lengthening has been observed in response to loss of fluid flow in kidney epithelial cells 31,32,18 and cytoskeletal disruption in human embryonic kidney cells. 22 Previous studies from our lab have shown that fluid flow in tendons occurs in response to tensile loading 25,27 and SD of tendon cells results in an immediate disruption of the actin cytoskeleton in these cells. 23 While the presence or absence of physical signals appears to be related to changes in cilia length, the precise mechanism(s) and pathways involved in mediating these changes are unknown.…”

Section: Discussionmentioning

confidence: 99%

“…9 While the precise mechanism(s) which trigger these changes in cilia length are unknown, it has been suggested that fluid flow and cell deformation are associated with decreases in cilia length, 9,[19][20][21] while increases in cilia length have been associated with cytoskeletal disruption. 22 Studies from our lab have shown that stress-deprivation (SD) of tendon cells has been associated with a disruption of cytoskeletal organization, 23 while tensile loading of tendons has been associated with cell deformation 24 and extracellular fluid flow. 25,26 Therefore, the purpose of the current study was to examine the effect of loading conditions on the length of primary cilia of rat tail tendon cells in situ.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Effect of in vitro stress‐deprivation and cyclic loading on the length of tendon cell cilia in situ

Gardner

Arnoczky

Lavagnino

2010

Journal Orthopaedic Research

View full text Add to dashboard Cite

To determine the effect of loading conditions on the length of primary cilia in tendon cells in situ, freshly harvested rat tail tendons were stress-deprived (SD) for up to 72 h, cyclically loaded at 3% strain at 0.17 Hz for 24 h, or SD for 24 h followed by cyclic loading (CL) for 24 h. Tendon sections were stained for tubulin, and cilia measured microscopically. In fresh control tendons, cilia length ranged from 0.6 to 2.0 mm with a mean length of 1.1 mm. Following SD, cilia demonstrated an increase ( p < 0.001) in overall length at 24 h when compared to controls. Cilia length did not increase with time of SD ( p ¼ 0.329). Cilia in cyclically loaded tendons were shorter ( p < 0.001) compared to all SD time periods, but were not different from 0 time controls ( p ¼ 0.472). CL for 24 h decreased cilia length in 24 h SD tendons ( p < 0.001) to levels similar to those of fresh controls ( p ¼ 0.274). The results of this study demonstrate that SD resulted in an immediate and significant increase in the length of primary cilia of tendon cells, which can be reversed by cyclic tensile loading. This suggests that, as in other tissues, cilia length in tendon cells is affected by mechanical signaling from the extracellular matrix. Keywords: tendon cell; primary cilia; stress-deprivation; tensile loading Mechanical loading is essential for maintaining the health and homeostasis of tendons, although the fundamental mechanotransduction pathways by which cells mediate this process are unclear. 1 Primary cilia are sensory organelles for the detection and transmission of mechanical and chemical information from the extracellular environment of cells [2][3][4] and are known to function as mechanosensors in several connective tissue cell types, including tenocytes, osteocytes, and chondrocytes. [5][6][7][8][9][10][11] The expression of transmembrane extracellular matrix receptors (integrins) on the cilium strongly suggests that the cilium has the molecular machinery necessary to act as a mechanosensory linkage between the ECM and cytoplasmic organelles. 10 Bending or stretching of the cilium can tug on such integrins, thus generating a cascade of events ranging from internal calcium release 12 to the activation of genes 10 and signaling molecules. [13][14][15] However, the degree of cilia bending required to elicit a specific cellular response is unknown. 16 The primary cilium has been modeled as a cantilevered beam whose deflection, secondary to cell deformation or fluid flow, produces a mechanosensory signal to the cell. 17 As such, is it logical to assume that the longer the beam (cilium), the more sensitive it will be to smaller deflection forces and, conversely, the shorter the beam, the less sensitive it will be. 17 Indeed, previous studies have suggested that the level of mechanical stimuli experienced by a cell could lead to remodeling of its ciliary structure through changes in its length. 16 Such modifications could, theoretically, make the cilia more or less sensitive to mechanical signals transmitted by the e...

show abstract

Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Effect of in vitro stress‐deprivation and cyclic loading on the length of tendon cell cilia in situ

Gardner

Arnoczky

Lavagnino

2010

Journal Orthopaedic Research

View full text Add to dashboard Cite

show abstract

“…13 Consistent with these observations, nucleating factors that promote actin polymerization repress primary cilium assembly and growth whereas actin severing enzymes promote ciliogenesis. [14][15][16][17] Actin depolymerization promotes ciliogenesis by stabilizing the pericentrosomal preciliary compartment, a vesiculotubular structure that contributes to ciliogenesis, 14,15,17 and by inhibiting transcription factors from the Hippo pathway, which induce the expression of cilium disassembly factors. 17 The small GTPase RhoA plays a key role in actin network regulation.…”

Section: Introductionmentioning

confidence: 99%

STAR syndrome-associated CDK10/Cyclin M regulates actin network architecture and ciliogenesis

et al. 2016

View full text Add to dashboard Cite

CDK10/CycM is a protein kinase deficient in STAR (toe Syndactyly, Telecanthus and Anogenital and Renal malformations) syndrome, which results from mutations in the X-linked FAM58A gene encoding Cyclin M. The biological functions of CDK10/CycM and etiology of STAR syndrome are poorly understood. Here, we report that deficiency of CDK10/Cyclin M promotes assembly and elongation of primary cilia. We establish that this reflects a key role for CDK10/Cyclin M in regulation of actin network organization, which is known to govern ciliogenesis. In an unbiased screen, we identified the RhoA-associated kinase PKN2 as a CDK10/ CycM phosphorylation substrate. We establish that PKN2 is a bone fide regulator of ciliogenesis, acting in a similar manner to CDK10/CycM. We discovered that CDK10/Cyclin M binds and phosphorylates PKN2 on threonines 121 and 124, within PKN2 0 s core RhoA-binding domain. Furthermore, we demonstrate that deficiencies in CDK10/CycM or PKN2, or expression of a non-phosphorylatable version of PKN2, destabilize both the RhoA protein and the actin network architecture. Importantly, we established that ectopic expression of RhoA is sufficient to override the induction of ciliogenesis resulting from CDK10/CycM knockdown, indicating that RhoA regulation is critical for CDK10/CycM's negative effect on ciliogenesis. Finally, we show that kidney sections from a STAR patient display dilated renal tubules and abnormal, elongated cilia. Altogether, these results reveal CDK10/CycM as a key regulator of actin dynamics and a suppressor of ciliogenesis through phosphorylation of PKN2 and promotion of RhoA signaling. Moreover, they suggest that STAR syndrome is a ciliopathy.

show abstract

“…Pitaval and colleagues (2010) found that actin cytoskeleton function was required for the response of cilia to the stretched area, which is consistent with the idea that actin is an integrator of global cellular mechanics. Actin has been implicated in ciliogenesis in large-scale screens (Kim et al 2010) although the mechanistic role of actin in ciliogenesis remains completely unknown.…”

Section: Mechanical Forces Affect Ciliogenesismentioning

confidence: 99%

Mechanobiology of Ciliogenesis

Ishikawa¹,

Marshall²

2014

BioScience

View full text Add to dashboard Cite

Cilia are force-generating and -sensing organelles that serve as mechanical interfaces between the cell and the extracellular environment. Cilia are present in tissues that adaptively respond to mechanical loading and fluid flow, and defects in ciliary function can lead to diseases affecting these tissues. As might be expected for a mechanical interface, the formation of cilia is, itself, regulated by mechanical forces, and these links between mechanics and ciliary formation are providing new entry points for dissecting the regulatory pathways of ciliogenesis.

show abstract

Functional genomic screen for modulators of ciliogenesis and cilium length

Cited by 487 publications

References 24 publications

Effect of in vitro stress‐deprivation and cyclic loading on the length of tendon cell cilia in situ

Effect of in vitro stress‐deprivation and cyclic loading on the length of tendon cell cilia in situ

STAR syndrome-associated CDK10/Cyclin M regulates actin network architecture and ciliogenesis

Mechanobiology of Ciliogenesis

Contact Info

Product

Resources

About