Nucleolar fibrillarin is an evolutionarily conserved regulator of bacterial pathogen resistance

Tiku, Varnesh; Kew, Chun; Mehrotra, Parul; Ganesan, Raja; Robinson, Nirmal; Antebi, Adam

doi:10.1038/s41467-018-06051-1

Cited by 48 publications

(83 citation statements)

References 47 publications

(54 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Puromycin incorporation was measured as previously described (Tiku et al, 2018) with minor modifications. Heat-inactivated OP-50 (75°C, 40 min) were provided as food source during pulse labelling.…”

Section: Methodsmentioning

confidence: 99%

The ribosomal RNA m⁵C methyltransferase NSUN-1 modulates healthspan and oogenesis inCaenorhabditis elegans

Heissenberger

Krammer

Rollins

et al. 2020

Preprint

View full text Add to dashboard Cite

21Our knowledge about the repertoire of ribosomal RNA modifications and the enzymes 22 responsible for installing them is constantly expanding. Previously, we reported that NSUN-5 23 is responsible for depositing m 5 C at position C2381 on the 26S rRNA in Caenorhabditis 24 elegans. 25 Here, we show that NSUN-1 is writing the second known 26S rRNA m 5 C at position C2982. 26 Depletion of nsun-1 or nsun-5 improved locomotion at midlife and resistance against heat 27 stress, however, only soma-specific knockdown of nsun-1 extended lifespan. Moreover, 28 soma-specific knockdown of nsun-1 reduced body size and impaired fecundity, suggesting 29 non-cell-autonomous effects. While ribosome biogenesis and global protein synthesis were 30 unaffected by nsun-1 depletion, translation of specific mRNAs was remodelled leading to 31 reduced production of collagens, loss of structural integrity of the cuticle and impaired barrier 32 function. 33 We conclude that loss of a single enzyme required for rRNA methylation has profound and 34 highly specific effects on organismal physiology. 102Therefore, we investigated the RNA substrate of NSUN-1 (also formerly known as NOL-1, 103 NOL-2 or W07E6.1) and its potential roles in worm physiology. NSUN-1 is also a member 104 of the NOP2/Sun RNA methyltransferase family. Since there are only two known m 5 C 105 residues on worm 26S rRNA (Sharma and Lafontaine, 2015; Trixl and Lusser, 2019), one of 106 them at C2381, being installed by NSUN-5, we speculated that NSUN-1 might be required 107 6 for introducing the second m 5 C residue at position C2982. Notably, both 26S m 5 C sites are 108 localized close to important functional regions of the ribosome, and they are highly conserved 109 during evolution ( Fig. 1A,B). 110In order to test if NSUN-1 is involved in large ribosomal subunit m 5 C methylation, 26S 111 rRNA was purified from worms treated with siRNAs specific to NSUN-1-encoding mRNAs 112 on sucrose gradients, digested to single nucleosides and analysed by quantitative HPLC. In 113 our HPLC assay, the m 5 C nucleoside eluted at 12 min, as established with a m 5 C calibration 114 control (data not shown). 115The depletion of NSUN-1 was conducted in two genetic backgrounds: N2 (wildtype), and 116 NL2099 (an RNAi-hypersensitive strain due to mutation in rrf-3) (Figure 1-figure 117 supplement 1). Treating N2 worms with an empty control vector, not expressing any RNAi, 118did not significantly reduce the levels of 26S rRNA m 5 C methylation (Fig. 1C, 97% instead 119 of 100%). Interestingly, treating N2 worms with an RNAi construct targeting nsun-1 led to a 120 reduction of 26S rRNA m 5 C methylation by 35% (Fig. 1C). In the NL2099 strain, nsun-1 121 RNAi treatment also led to a reduction of 26S rRNA m 5 C methylation by 26% ( Fig. 1D). 122As there are only two known modified m 5 C residues on worm 26S rRNA, and since one of 123 them is introduced by NSUN-5 (Adamla et al., 2019; Schosserer et al., 2015), a complete loss 124 of NSUN-1 activity was expected to result in a 50% decrease...

show abstract

Section: Methodsmentioning

confidence: 99%

The ribosomal RNA m⁵C methyltransferase NSUN-1 modulates healthspan and oogenesis inCaenorhabditis elegans

Heissenberger

Krammer

Rollins

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…23 Young adult worms were transferred to killing plate (tryptic soy agar/TSA medium with S. aureus strain HG001, or NGM containing 0.35% peptone with P. aeruginosa strain PA14) and scored every 12 hours at 25°C. 23 Young adult worms were transferred to killing plate (tryptic soy agar/TSA medium with S. aureus strain HG001, or NGM containing 0.35% peptone with P. aeruginosa strain PA14) and scored every 12 hours at 25°C.…”

Section: Elegans Killing Assaymentioning

confidence: 99%

“…23 Human HeLa cells with USP36 overexpression or knockdown were infected by S. aureus (MOI 50) for the indicated time points. 23 Human HeLa cells with USP36 overexpression or knockdown were infected by S. aureus (MOI 50) for the indicated time points.…”

Section: Bacteria Killing and Clearance Assaysmentioning

confidence: 99%

“…Recently, Tiku and colleagues demonstrated that upon bacterial infection worms downregulate fibrillarin as an antipathogen protective response 23 ; however, the mechanism underlying this fibrillarin reduction remains unresolved. Based on the fibrillarin-targeting regulatory pathway delineated by our study thus far, we thus postulated that CEP-1, CED-3, and FIB-1 would be modulated coordinately when worms are challenged with bacteria.…”

Section: The Cep-1-ced-3-usp-33 Axis Protects Against Bacterial Infmentioning

confidence: 99%

“…15,16 The tumorigenic link of the nucleolus is further evidenced by the functional interaction with tumor suppressor proteins. 23 However, how the host cells transduce this nucleolus-associated anti-bacterial signaling remains largely unresolved. 17 The notion that the nucleolus acts as intracellular stress sensor is also gaining ground, as its alteration is linked to a multitude of "nucleolar stresses," including DNA damage, impaired RNA and protein synthesis, nutrient starvation, heat or cold shock, hypoxia, and oxidative stress.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Nucleolar control by a non‐apoptotic p53‐caspases‐deubiquitinylase axis promotes resistance to bacterial infection

Chen

Chu

et al. 2019

FASEB j.

View full text Add to dashboard Cite

This is an open access article under the terms of the Creat ive Commo ns Attri butio n-NonCo mmercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. AbstractThe nucleolus is best known for its cellular role in regulating ribosome production and growth. More recently, an unanticipated role for the nucleolus in innate immunity has recently emerged whereby downregulation of fibrillarin and nucleolar contraction confers pathogen resistance across taxa. The mechanism of this downregulation, however, remains obscure. Here we report that rather than fibrillarin itself being the proximal factor in this pathway, the key player is a fibrillarin-stabilizing deubiquitinylase USP-33. This was discovered by a candidate-gene search of Caenorhabditis elegans in which CED-3 caspase was revealed to execute targeted cleavage of USP-33, thus destabilizing fibrillarin. We also showed that cep-1 and ced-3 mutant worms altered nucleolar size and decreased antimicrobial peptide gene, spp-1, expression rendering susceptibility to bacterial infection. These phenotypes were reversed by usp-33 knockdown, thus linking the CEP-1-CED-3-USP-33 pathway with nucleolar control and resistance to bacterial infection in worms. Parallel experiments with the human analogs of caspases and USP36 revealed similar roles in coordinating these two processes. In summary, our work outlined a conserved cascade that connects cell death signaling to nucleolar control and innate immune response.

show abstract

C. elegans: out on an evolutionary limb

Pujol

Ewbank

2021

Immunogenetics

View full text Add to dashboard Cite

The natural environment of the free-living nematode Caenorhabditis elegans is rich in pathogenic microbes. There is now ample evidence to indicate that these pathogens exert a strong selection pressure on C. elegans, and have shaped its genome, physiology, and behaviour. In this short review, we concentrate on how C. elegans stands out from other animals in terms of its immune repertoire and innate immune signalling pathways. We discuss how C. elegans often detects pathogens because of their effects on essential cellular processes, or organelle integrity, in addition to direct microbial recognition. We illustrate the extensive molecular plasticity that is characteristic of immune defences in C. elegans and highlight some remarkable instances of lineage-specific innovation in innate immune mechanisms.

show abstract

Nucleolar fibrillarin is an evolutionarily conserved regulator of bacterial pathogen resistance

Cited by 48 publications

References 47 publications

The ribosomal RNA m⁵C methyltransferase NSUN-1 modulates healthspan and oogenesis inCaenorhabditis elegans

The ribosomal RNA m⁵C methyltransferase NSUN-1 modulates healthspan and oogenesis inCaenorhabditis elegans

Nucleolar control by a non‐apoptotic p53‐caspases‐deubiquitinylase axis promotes resistance to bacterial infection

C. elegans: out on an evolutionary limb

Contact Info

Product

Resources

About

Nucleolar fibrillarin is an evolutionarily conserved regulator of bacterial pathogen resistance

Cited by 48 publications

References 47 publications

The ribosomal RNA m5C methyltransferase NSUN-1 modulates healthspan and oogenesis inCaenorhabditis elegans

The ribosomal RNA m5C methyltransferase NSUN-1 modulates healthspan and oogenesis inCaenorhabditis elegans

Nucleolar control by a non‐apoptotic p53‐caspases‐deubiquitinylase axis promotes resistance to bacterial infection

C. elegans: out on an evolutionary limb

Contact Info

Product

Resources

About

The ribosomal RNA m⁵C methyltransferase NSUN-1 modulates healthspan and oogenesis inCaenorhabditis elegans

The ribosomal RNA m⁵C methyltransferase NSUN-1 modulates healthspan and oogenesis inCaenorhabditis elegans