P granules extend the nuclear pore complex environment in the <i>C. elegans</i> germ line

Updike, Dustin L.; Hachey, Stephanie J.; Kreher, Jeremy; Strome, Susan

doi:10.1083/jcb.201010104

Cited by 227 publications

(249 citation statements)

References 44 publications

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“…Hexanediol was shown to perturb FG repeat interactions between nucleoporins in nuclear pores [6] [7] and interactions between RNAbinding proteins in RNA-protein granules (RNP) [2] [8]. These studies suggested that 1,6-hexanediol interferes with weak hydrophobic protein-protein or protein-RNA interactions that are required for these dynamic, liquid-like assemblies to form.…”

Section: Resultsmentioning

confidence: 99%

Hexanediol: a chemical probe to investigate the material properties of membrane-less compartments

Kroschwald

Maharana

Alberti³

2017

Matters

293

277

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Membrane-less compartments, such as P bodies and stress granules, play major roles in cell biology and disease. The dynamics and physical properties of these compartments are very diverse, ranging from liquid-like to solid-like behavior. Importantly, changes in compartment dynamics have been linked to changes in compartment function or pathology. However, appropriate tools to investigate the physical properties of membrane-less compartments are still lacking. The aliphatic alcohol hexanediol has been proposed as a tool to differentiate between liquid-like and solid-like assemblies in living cells. We show that in vitro reconstituted compartments formed by the RNAbinding protein FUS are rapidly dissolved by hexanediol. In contrast, solid-like fibers of FUS, which have been linked to the disease amyotrophic lateral sclerosis (ALS), are resistant to hexanediol treatment, supporting the idea that hexanediol can differentiate between liquid-and solid-like assemblies. We further show that hexanediol can be used to examine the physical properties of membrane-less compartments in vivo. We find that hexanediol dissolves dynamic, liquid-like assemblies, such as P bodies, whereas solid-like assemblies, such as protein aggregates and cytoskeletal assemblies, are largely resistant to hexanediol. Finally, we report here that extended exposure of yeast and mammalian cells to hexanediol is cytotoxic and causes abnormal changes in cell morphology, which trigger the formation of aberrant assemblies. We therefore urge care in the use of hexanediol, especially when cells are exposed to hexanediol for extended times. In summary, we propose that hexanediol is a powerful tool to probe the physical properties of membrane-less compartments, but only if adequate controls are included and precautions are taken.

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Section: Resultsmentioning

confidence: 99%

Hexanediol: a chemical probe to investigate the material properties of membrane-less compartments

Kroschwald

Maharana

Alberti³

2017

Matters

293

277

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“…Second, because phase separated droplets are >90 % water by mass (based on protein concentrations ~300 µM), macromolecules could enter, diffuse within and exit easily. Indeed, 40 kDa dextrans readily enter P-granules in nematodes, although 155 kD dextrans are excluded (Updike et al, 2011). Together, these properties conspire to enable molecules to be concentrated within granules, but move rapidly and freely within them.…”

Section: Discussionmentioning

confidence: 99%

“…These sequences were originally identified by their similarity to those of the known prions, human prion protein and Sup35p (Gilks et al, 2004; Riejns et al, 2008; Decker et al, 2007), but also include those containing repeated G/S-F/Y-G/S motifs (Kato et al, 2012; Nott et al, 2015; Updike et al, 2011; King et al, 2012; Sun et al, 2011). These sequences can be important for targeting to, and/or formation of, RNP granules.…”

Section: Introductionmentioning

confidence: 99%

“…Similarly, P-bodies in yeast assemble through redundant interactions of the Edc3 protein and by a prion domain of Lsm4 (Decker et al, 2007), while an mRNP granule in the fungus Ashbya is driven by a polyQ region in the Whi3 protein (Lee et al, 2013). The P-granules in C. elegans are dependent on the Pgl family of proteins for their assembly, which contain an XFG repeat structure (Updike et al, 2011). In addition, the RNA binding protein Fus localizes to yeast and mammalian stress granules through its N-terminal IDR, and mutation of tyrosines in multiple GYG motifs in this region prevents this accumulation (Kato et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

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Formation and Maturation of Phase-Separated Liquid Droplets by RNA-Binding Proteins

et al. 2015

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Eukaryotic cells possess numerous dynamic membrane-less organelles, RNP granules, enriched in RNA and RNA binding proteins containing disordered regions. We demonstrate that the disordered regions of key RNP granule components, and the full-length granule protein hnRNPA1, can phase separate in vitro, producing dynamic liquid droplets. Phase separation is promoted by low salt concentrations or RNA. Over time, the droplets mature to more stable states, as assessed by slowed fluorescence recovery after photobleaching and resistance to salt. Maturation often coincides with formation of fibrous structures. Different disordered domains can co-assemble into phase-separated droplets. These biophysical properties demonstrate a plausible mechanism by which interactions between disordered regions, coupled with RNA binding, could contribute to RNP granule assembly in vivo through promoting phase separation. Progression from dynamic liquids to stable fibers may be regulated to produce cellular structures with diverse physiochemical properties and functions. Misregulation could contribute to diseases involving aberrant RNA granules.

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“…Furthermore, FG repeats, a hallmark of many nuclear pore proteins, have been found in another family of P-granule proteins comprising GLH-1, -2 and -4 (Kuznicki et al, 2000). Based on these findings, P granules are considered as extensions of nuclear pore complexes (Updike et al, 2011). P granules might confer germline-specific roles on nuclear pore complexes.…”

Section: Discussionmentioning

confidence: 99%

Loss of PGL-1 and PGL-3, a family of constitutive germ-granule components, promotes germline apoptosis in C. elegans

Min

Shim

Kawasaki

2015

Journal of Cell Science

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In Caenorhabditis elegans, the mechanisms regulating germline apoptosis remain largely unknown, except for the core machinery. Here, we found that mutants of pgl-1 and pgl-3, encoding members of a family of constitutive protein components of germline-specific P granules, showed increased germline apoptosis under both physiological and DNA-damaged conditions. We also found that the number of germ cells that lost PGL proteins increased significantly following UV irradiation, and that only those PGL-absent germ cells were selectively engulfed by gonadal sheath cells in adult hermaphrodite gonads. We further revealed that CEP-1, the p53 homolog, and the caspase CED-3 promoted elimination of PGL-1 from germ cells following UV irradiation. Furthermore, protein levels of CED-4, the Apaf-1 homolog, and cytoplasmic translocation of SIR-2.1, a Sirtuin homolog, significantly increased in pgl mutants and increased even more following UV irradiation. CED-4 and SIR-2.1 were essential for high levels of germline apoptosis in pgl mutants. We conclude that PGL proteins suppress excessive germline apoptosis by repressing both the protein levels of CED-4 and the cytoplasmic translocation of SIR-2.1. Our study has revealed new roles for PGL-1 and PGL-3 in the control of germline apoptosis.

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P granules extend the nuclear pore complex environment in the C. elegans germ line

Abstract: Like the nuclear pore complex, FG repeat–containing P-granule proteins interact to help establish a size-exclusion barrier.

Cited by 227 publications

References 44 publications

Hexanediol: a chemical probe to investigate the material properties of membrane-less compartments

Hexanediol: a chemical probe to investigate the material properties of membrane-less compartments

Formation and Maturation of Phase-Separated Liquid Droplets by RNA-Binding Proteins

Loss of PGL-1 and PGL-3, a family of constitutive germ-granule components, promotes germline apoptosis in C. elegans

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