ZYX-1/Zyxin plays a minor role in oocyte transit through the spermatheca in C. elegans

Castaneda, Perla G.; Wu, Nan; Qiu, Zhongqiang; Lee, Myeongwoo; Cram, Erin J.

doi:10.17912/micropub.biology.000489

Cited by 3 publications

(3 citation statements)

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“…4B and C). In young adults we could see mNG::ZYX-1a readily in differentiated body wall muscle, neurons, gonads, and spermatheca, in line with where its expression was previously described (46, 54) (Supplementary Fig. 4D).…”

Section: Resultssupporting

confidence: 90%

Zyxin contributes to coupling between cell junctions and contractile actomyosin networks during apical constriction

Slabodnick

Tintori

Prakash

et al. 2022

Preprint

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One of the most common cell shape changes driving morphogenesis in diverse animals is the constriction of the apical cell surface. Apical constriction depends on contraction of an actomyosin network in the apical cell cortex, but such actomyosin networks have been shown to undergo continual, conveyor belt-like contractions even before the shrinking of an apical surface begins. This finding suggests that apical constriction is not necessarily triggered by the contraction of actomyosin networks, but rather by unidentified, temporally-regulated mechanical links between actomyosin and junctions. Here, we used C. elegans gastrulation as a model to identify proteins that contribute to such linkage. We found that α-catenin and β-catenin initially failed to move centripetally with contracting cortical actomyosin networks, suggesting that linkage is regulated between cadherin-catenin complexes and actomyosin. We used a proteomic approach to identify AFD-1/afadin and transcriptomics to identify ZYX-1/zyxin as contributors to C. elegans gastrulation. We found that a zyxin family of LIM domain proteins have transcripts that become enriched in multiple cells just before they undergo apical constriction. Using a new, semi-automated image analysis tool, we found that AFD-1/afadin and ZYX-1/zyxin both contribute to cell-cell junctions’ centripetal movement in concert with contracting actomyosin networks. These results identify two key proteins as important for actomyosin networks to effectively pull cell-cell junctions inward during apical constriction. The transcriptional upregulation of zyxin/ZYX-1 in specific cells points to one way that developmental patterning spatiotemporally regulates cell biological mechanisms in vivo. Because afadin- and zyxin-family proteins contribute to membrane-cytoskeleton linkage in other systems, we anticipate that their roles in regulating apical constriction in this manner may be conserved.

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Section: Resultssupporting

confidence: 90%

Zyxin contributes to coupling between cell junctions and contractile actomyosin networks during apical constriction

Slabodnick

Tintori

Prakash

et al. 2022

Preprint

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“…Previous authors have reported that zyx-1/zyxin produces 2 protein isoforms: a longer, 603 amino acid isoform called ZYX-1a, and a shorter, 200 amino acid isoform called ZYX-1b [46]. ZYX-1a contains 3 polyproline-rich repeats, a predicted nuclear export signal, and 3 tandem LIM domains (S4A Fig) . We created a strain with mNeonGreen (mNG) inserted at the endogenous N-terminus to tag ZYX-1a, but consistent with its low predicted expression at this stage of development, we were unable to detect mNG signal in the EPCs, and we were unable to detect ZYX-1 by immunostaining methods that employed signal amplification (S4B and S4C Fig) . In young adults we could see mNG::ZYX-1a readily in differentiated body wall muscle, neurons, gonads, and spermatheca, in line with where its expression was previously described [46,54] (S4D Fig) . To assess where ZYX-1 could associate when accumulating in gastrulating cells, we examined where overexpressed ZYX-1 would localize using single-copy transgenes driven by the sdz-1 promoter (Psdz-1), which is predicted to drive ~20-fold overexpression compared to zyx-1 expression levels in EMS, E, and MS cell lineages (S5A Fig) . We created two mNG-tagged constructs: one expressing full length ZYX-1a, and another expressing only the LIM domain-containing region (LCR) of ZYX-1 to examine where the LCR alone could direct localization. For both constructs, cytoplasmic mNG signal could be detected in E and MS cells as predicted, and small foci could be detected at the apical surfaces of internalizing EPCs (S5B Fig) . Additionally, the predicted nuclear export signal of ZYX-1a appeared to be functional in full-length mNG::ZYX-1a, because mNG::ZYX-1a was excluded from the nucleus while mNG::LCR ZYX-1 was not (S5C Fig) . We conclude that ZYX-1a is likely expressed normally at too low a level as EPCs internalize to detect by current methods, and that it and its LCR can be recruited to apical foci in EPCs when overexpressed.…”

Section: Plos Geneticssupporting

confidence: 89%

Zyxin contributes to coupling between cell junctions and contractile actomyosin networks during apical constriction

et al. 2023

View full text Add to dashboard Cite

One of the most common cell shape changes driving morphogenesis in diverse animals is the constriction of the apical cell surface. Apical constriction depends on contraction of an actomyosin network in the apical cell cortex, but such actomyosin networks have been shown to undergo continual, conveyor belt-like contractions before the shrinking of an apical surface begins. This finding suggests that apical constriction is not necessarily triggered by the contraction of actomyosin networks, but rather can be triggered by unidentified, temporally-regulated mechanical links between actomyosin and junctions. Here, we used C. elegans gastrulation as a model to seek genes that contribute to such dynamic linkage. We found that α-catenin and β-catenin initially failed to move centripetally with contracting cortical actomyosin networks, suggesting that linkage is regulated between intact cadherin-catenin complexes and actomyosin. We used proteomic and transcriptomic approaches to identify new players, including the candidate linkers AFD-1/afadin and ZYX-1/zyxin, as contributing to C. elegans gastrulation. We found that ZYX-1/zyxin is among a family of LIM domain proteins that have transcripts that become enriched in multiple cells just before they undergo apical constriction. We developed a semi-automated image analysis tool and used it to find that ZYX-1/zyxin contributes to cell-cell junctions’ centripetal movement in concert with contracting actomyosin networks. These results identify several new genes that contribute to C. elegans gastrulation, and they identify zyxin as a key protein important for actomyosin networks to effectively pull cell-cell junctions inward during apical constriction. The transcriptional upregulation of ZYX-1/zyxin in specific cells in C. elegans points to one way that developmental patterning spatiotemporally regulates cell biological mechanisms in vivo. Because zyxin and related proteins contribute to membrane-cytoskeleton linkage in other systems, we anticipate that its roles in regulating apical constriction in this manner may be conserved.

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“…There are anatomical differences in isoform expression, with ZYX-1b observed in body wall muscle, pharynx, vulva, spermathecae, and multiple neurons, whereas ZYX-1a mainly localizes to tail phasmid neurons and uterine muscle, and weakly so, body wall muscle [13]. In C. elegans, zyx-1 has been postulated to have a minor role in reproduction, however, the mechanism(s) and isoform (s) involved remain elusive [14,15]. Beyond this, C. elegans zyx-1 is hypothesized to be functionally analogous to vertebrate zyxin, with LIM domains acting as mechanical stabilizers at focal adhesions, and the proline-rich N-terminus involved in sensing muscle cell damage [12].…”

Section: Introductionmentioning

confidence: 99%

azyx-1 is a new gene that overlaps with zyxin and affects its translation in C. elegans, impacting muscular integrity and locomotion

Parmar,

Kieswetter,

Geens

et al. 2023

PLoS Biol

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Overlapping genes are widely prevalent; however, their expression and consequences are poorly understood. Here, we describe and functionally characterize a novel zyx-1 overlapping gene, azyx-1, with distinct regulatory functions in Caenorhabditis elegans. We observed conservation of alternative open reading frames (ORFs) overlapping the 5′ region of zyxin family members in several animal species, and find shared sites of azyx-1 and zyxin proteoform expression in C. elegans. In line with a standard ribosome scanning model, our results support cis regulation of zyx-1 long isoform(s) by upstream initiating azyx-1a. Moreover, we report on a rare observation of trans regulation of zyx-1 by azyx-1, with evidence of increased ZYX-1 upon azyx-1 overexpression. Our results suggest a dual role for azyx-1 in influencing zyx-1 proteoform heterogeneity and highlight its impact on C. elegans muscular integrity and locomotion.

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ZYX-1/Zyxin plays a minor role in oocyte transit through the spermatheca in C. elegans

Cited by 3 publications

References 0 publications

Zyxin contributes to coupling between cell junctions and contractile actomyosin networks during apical constriction

Zyxin contributes to coupling between cell junctions and contractile actomyosin networks during apical constriction

Zyxin contributes to coupling between cell junctions and contractile actomyosin networks during apical constriction

azyx-1 is a new gene that overlaps with zyxin and affects its translation in C. elegans, impacting muscular integrity and locomotion

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