Lambacher NJ et al. (2016) Nat Cell Biol "TMEM107 recruits ciliopathy proteins to subdomains of the ciliary transition zone and causes Joubertsyndrome."

Riviere JB, Kuhns S, Giles RH, Lambacher NJ, van Dam TJ, Szymaska K, McManus GJ, Peckham M, Curd A, Saunier S, Wu KM, Attie-Bitach T, Thauvin-Robinet C, van der Lee R, Doummar D, Faivre L, Johnson CA, Slaats GG, Burglen L, Gaff K, Kennedy JE, Bruel AL, Huynen MA, Blacque OE

[Nat Cell Biol, 2016]

The transition zone (TZ) ciliary subcompartment is thought to control cilium composition and signalling by facilitating a protein diffusion barrier at the ciliary base. TZ defects cause ciliopathies such as Meckel-Gruber syndrome (MKS), nephronophthisis (NPHP) and Joubert syndrome (JBTS). However, the molecular composition and mechanisms underpinning TZ organization and barrier regulation are poorly understood. To uncover candidate TZ genes, we employed bioinformatics (coexpression and co-evolution) and identified TMEM107 as a TZ protein mutated in oral-facial-digital syndrome and JBTS patients. Mechanistic studies in Caenorhabditis elegans showed that TMEM-107 controls ciliary composition and functions redundantly with NPHP-4 to regulate cilium integrity, TZ docking and assembly of membrane to microtubule Y-link connectors. Furthermore, nematode TMEM-107 occupies an intermediate layer of the TZ-localized MKS module by organizing recruitment of the ciliopathy proteins MKS-1, TMEM-231 (JBTS20) and JBTS-14 (TMEM237). Finally, MKS module membrane proteins are immobile and super-resolution microscopy in worms and mammalian cells reveals periodic localizations within the TZ. This work expands the MKS module of ciliopathy-causing TZ proteins associated with diffusion barrier formation and provides insight into TZ subdomain architecture.

Musset-Bilal F et al. (1994) Worm Breeder's Gazette "Characterization of the C. elegans Basement Membrane-Associated Protein SPARC"

Musset-Bilal F, Schwarzbauer JE, Ryan CS

[Worm Breeder's Gazette, 1994]

Characterization of the C. elegans Basement Membrane- Associated Frederique Musset-Bilal, Carol S. Ryan, and Jean E. Schwarzbauer Department of Molecular Biology, Princeton University, Princeton NJ 08544

Townsend SR et al. (1994) Worm Breeder's Gazette "C. elegans Molecular Genetics and Long PCR."

Finelli AL, Savage C, Xie T, Padgett RW, Townsend SR

[Worm Breeder's Gazette, 1994]

C. elegans Molecular Genetics and Long PCR Scott R. Townsend, Cathy Savage, Alyce L. Finelli, Ting Xie, and Richard W. Padgett, Waksman Institute and Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, NJ 08855

Wilkinson HA et al. (1994) Worm Breeder's Gazette "Reciprocal Changes in Expression of lin-12 and lag-2 Prior to Commitment of Z1.ppp and Z4.aaa."

Greenwald IS, Wilkinson HA, Fitzgerald K

[Worm Breeder's Gazette, 1994]

Reciprocal changes in expression of lin-12 and lag-2 prior to commitment of Z1.ppp and Z4.aaa. Hilary A. Wilkinson*, Kevin Fitzgerald and Iva Greenwald, HHMI and Dept. of Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York, NY 10032; *Current address: Merck Research Laboratories, Rahway, NJ 07065

Ishii N et al. (1988) Worm Breeder's Gazette "partial sequence of the unc-6 gene."

Ishii N, Stern BD, Culotti JG, Hedgecock EM

[Worm Breeder's Gazette, 1988]

In the previous issue (WBG 10(1) p.35), we reported cloning a small DNA fragment containing the Tc1 insertion which caused the unc-6 ( rh1035) mutation. Since then, we have isolated an 11.5 kb EcoRI fragment including this region from N2 DNA (NJ#14, EMBL4 vector) Alan Coulson has kindly positioned this fragment on the MRC genome map. We have now sequenced both the original 1.25 kb XbaI fragment of unc-6( rh1035) DNA (plasmid NJ#6) and a 2.7 kb HindIII fragment of N2 DNA subcloned from phage NJ#14. The 2.7 kb HindIII fragment has five exons which encode a hypothetical protein fragment of 360 residues. This fragment is not closely related to any protein in the current PIR database The last exon in the 2.7 kb HindIII fragment is followed by an intron which continues beyond the fragment. The first exon is preceded by a non-coding region which may be part of an intron or the unc-6 promoter region. In fact, a possible 21 residue signal sequence with an initiator methionine occurs in the first exon shortly upstream of the Tc1::rh1035 insertion site. TATA and CCAAT motifs occur about 90 and 120 bp upstream of the proposed initiator codon. The decanucleotide sequence TTTGCCTCTC is tandemly repeated about 200 bp upstream of this codon. [See Figure 1]

Padmanaban S et al. (2024) Proc Natl Acad Sci U S A "Caenorhabditis elegans telomere-binding proteins TEBP-1 and TEBP-2 adapt the Myb module to dimerize and bind telomeric DNA."

Lambacher NJ, Nandakumar J, Shibuya H, Padmanaban S, Zhang J, Tesmer VM

[Proc Natl Acad Sci U S A, 2024]

Protecting chromosome ends from misrecognition as double-stranded (ds) DNA breaks is fundamental to eukaryotic viability. The protein complex shelterin prevents a DNA damage response at mammalian telomeres. Mammalian shelterin proteins TRF1 and TRF2 and their homologs in yeast and protozoa protect telomeric dsDNA. N-terminal homodimerization and C-terminal Myb-domain-mediated dsDNA binding are two structural hallmarks of end protection by TRF homologs. Yet our understanding of how <i>Caenorhabditis elegans</i> protects its telomeric dsDNA is limited. Recently identified <i>C. elegans</i> proteins TEBP-1 (also called DTN-1) and TEBP-2 (also called DTN-2) are functional homologs of TRF proteins, but how they bind DNA and whether or how they dimerize is not known. TEBP-1 and TEBP-2 harbor three Myb-containing domains (MCDs) and no obvious dimerization domain. We demonstrate biochemically that only the third MCD binds DNA. We solve the X-ray crystal structure of TEBP-2 MCD3 with telomeric dsDNA to reveal the structural mechanism of telomeric dsDNA protection in <i>C. elegans</i>. Mutagenesis of the DNA-binding site of TEBP-1 and TEBP-2 compromises DNA binding in vitro, and increases DNA damage signaling, lengthens telomeres, and decreases brood size in vivo. Via an X-ray crystal structure, biochemical validation of the dimerization interface, and SEC-MALS analysis, we demonstrate that MCD1 and MCD2 form a composite dimerization module that facilitates not only TEBP-1 and TEBP-2 homodimerization but also heterodimerization. These findings provide fundamental insights into <i>C. elegans</i> telomeric dsDNA protection and highlight how different eukaryotes have evolved distinct strategies to solve the chromosome end protection problem.

Toth, Marton L. et al. (2009) International Worm Meeting "HSF-1 is an Essential Downstream Mediator of Resveratrol-induced, SIR-2.1-dependent Longevity in C. elegans."

Soti, Csaba, Toth, Marton L., Csermely, Peter, Dancso, Balazs

[International Worm Meeting, 2009]

Major longevity assurance mechanisms include the heat-shock response governed by the heat-shock transcription factor (HSF-1) and the metabolic stress response, exemplified by the Sir2 deacetylase. Sir2, and recently HSF-1 have been implicated in the life-span extending effect of dietary restriction. Resveratrol, a polyphenol from red wine, is a dietary restriction mimetic that induces longevity and displays an impressive therapeutic potential against various degenerative diseases via Sir2. Resveratrol has also been shown to induce the heat-shock response in mammalian cells. In this study, we have asked how the heat-shock response is involved in resveratrol-induced/Sir2-related longevity in C. elegans. We find that resveratrol specifically induces chaperone expression, thermotolerance and longevity depending on both SIR-2.1 and HSF-1. Moreover, we show that SIR-2.1 is a potent activator of HSF-1-dependent thermotolerance. Finally, we identify HSF-1 as an essential downstream effector of resveratrol-induced, SIR-2.1-dependent longevity in C. elegans. Thus, our results demonstrate a direct interaction of metabolic and proteotoxic stress responses in life-span regulation and indicate that the robustness of the heat-shock response may determine the therapeutic response to resveratrol. Marton Toth''s current address: Rutgers, The State University of NJ, Molecular Biology & Biochemistry, Piscataway, NJ.

Kazatskaya A et al. (2017) Genetics "Primary Cilium Formation and Ciliary Protein Trafficking Is Regulated by the Atypical MAP Kinase MAPK15 in Caenorhabditis elegansand Human Cells."

Brear AG, Blacque OE, Kuhns S, Kazatskaya A, Kennedy JE, Sengupta P, McManus GJ, Lambacher NJ

[Genetics, 2017]

Motile and immotile (or primary) cilia are microtubule-based structures that mediate multiple cellular functions including transduction of environmental cues, developmental signaling, cellular motility, and modulation of fluid flow. Although their core architectures are similar, motile and primary cilia exhibit marked structural differences that underlie distinct functional properties. However, the extent to which ciliogenesis mechanisms are shared between these different cilia types is not fully described. Here we report that the atypical MAP kinase MAPK15 (ERK7/8), implicated in the formation of vertebrate motile cilia, also regulates the formation of primary cilia in C. elegans sensory neurons and human cells. We find that MAPK15 localizes to a basal body subdomain with the ciliopathy protein BBS7 and cell-cell junctions. MAPK15 also regulates the localization of ciliary proteins involved in cilium structure, transport, and signaling. Our results describe a primary cilia-related role for this poorly studied member of the MAPK family in vivo, and indicate a broad requirement for MAPK15 in the formation of multiple ciliary classes across species.

Jensen VL et al. (2018) EMBO Rep "Role for intraflagellar transport in building a functional transition zone."

Jensen VL, Inglis PN, Lambacher NJ, Li C, Leroux MR, Blacque OE, Mohan S, Yoder BK, Williams CL

[EMBO Rep, 2018]

Genetic disorders caused by cilia dysfunction, termed ciliopathies, frequently involve the intraflagellar transport (IFT) system. Mutations in IFT subunits-including IFT-dynein motor DYNC2H1-impair ciliary structures and Hedgehog signalling, typically leading to &quot;skeletal&quot; ciliopathies such as Jeune asphyxiating thoracic dystrophy. Intriguingly, IFT gene mutations also cause eye, kidney and brain ciliopathies often linked to defects in the transition zone (TZ), a ciliary gate implicated in Hedgehog signalling. Here, we identify a <i>C.elegans</i> temperature-sensitive (<i>ts</i>) IFT-dynein mutant (<i>che-3</i>; human DYNC2H1) and use it to show a role for retrograde IFT in anterograde transport and ciliary maintenance. Unexpectedly, correct TZ assembly and gating function for periciliary proteins also require IFT-dynein. Using the reversibility of the novel <i>ts</i>-IFT-dynein, we show that restoring IFT in adults (post-developmentally) reverses defects in ciliary structure, TZ protein localisation and ciliary gating. Notably, this ability to reverse TZ defects declines as animals age. Together, our findings reveal a previously unknown role for IFT in TZ assembly in metazoans, providing new insights into the pathomechanism and potential phenotypic overlap between IFT- and TZ-associated ciliopathies.

Jenkins NL et al. (2000) European Worm Meeting "C. elegans and humans share similar patterns of non-linear growth"

Lithgow GJ, Clayton PE, Jenkins NL, Foster P, Gill MS

[European Worm Meeting, 2000]

Human growth is generally considered to be a relatively smooth, continuous process. However when examined frequently over short time periods growth is non-linear, with variations in height occurring at daily, weekly, monthly and annual intervals. This pattern of growth is likely to be dictated by nutritional, environmental, hormonal and genetic influences, but delineation of the relative contribution of each is not practical or ethical in humans. We have therefore sought to establish C. elegans as a model in which to characterise the processes that underpin non-linear growth. We have examined short-term growth by video microscopy in the Bristol N2 strain with size determinations made every 15 minutes over 12 hours in both L1 and L3. Length and area were calculated from triplicate images. Using curve fitting, individual growth curves exhibited periods of rapid growth interspersed by periods of little or no growth, consistent with the non-linear growth observed in human studies. Smoothed empirical growth velocities were calculated as the difference between successive measurements, scaled by the intervening time period. These data suggest that L1 growth can be characterised by 5-7 growth spurts of duration 80-95 min, with velocity ranging from a peak of 0.27-0.34m/min to a nadir of 0.01-0.05m/min. We are particularly interested in trying to relate the pattern of short-term growth to events in the cell lineage. Moreover the apparent conservation of short-term growth patterns between nematodes and humans suggests that C. elegans could be used to uncover novel genes that influence the kinetics of growth in both species. comments: Affiliations: Academic Unit of Child Health (MG, PC), School of Biological Sciences (MG, NJ, GL), Dept of Mathematics (PF).