The Arabidopsis XBAT35 is one of five structurally related ankyrin repeat-containing Really Interesting New Gene （RING） E3 ligases involved in ubiquitin-mediated protein degradation, which plays key roles in a wide ...The Arabidopsis XBAT35 is one of five structurally related ankyrin repeat-containing Really Interesting New Gene （RING） E3 ligases involved in ubiquitin-mediated protein degradation, which plays key roles in a wide range of cel- lular processes. Here, we show that the XBAT35 gene undergoes alternative splicing, generating two transcripts that are constitutively expressed in all plant tissues. The two splice variants derive from an exon skipping event that excludes an in-frame segment from the XBAT35 precursor mRNA, giving rise to two protein isoforms that differ solely in the presence of a nuclear localization signal （NLS）. Transient expression assays indicate that the isoform lacking the NLS localizes in the cytoplasm of plant cells, whereas the other is targeted to the nucleus, accumulating in nuclear speckles. Both isoforms are functional E3 ligases, as assessed by in vitro ubiquitination assays. Two insertion mutant alleles and RNA-interference （RNAi） silencing lines for XBAT35 display no evident phenotypes under normal growth conditions, but exhibit hyper- sensitivity to the ethylene precursor 1-aminocyclopropane-l-carboxylate （ACC） during apical hook exaggeration in the dark, which is rescued by an inhibitor of ethylene perception. Independent expression of each XBAT35 splice variant in the mutant background indicates that the two isoforms may differentially contribute to apical hook formation but are both functional in this ethylene-mediated response. Thus, XBAT35 defines a novel player in ethylene signaling involved in negatively regulating apical hook curvature, with alternative splicing controlling dual targeting of this E3 ubiquitin ligase to the nuclear and cytoplasmic compartments.展开更多
The development of a hook-like structure at the apical part of the soil-emerging organs has fascinated botanists for centuries,but how it is initiated remains unclear.Here,we demonstrate with highthroughput infrared i...The development of a hook-like structure at the apical part of the soil-emerging organs has fascinated botanists for centuries,but how it is initiated remains unclear.Here,we demonstrate with highthroughput infrared imaging and 2-D clinostat treatment that,when gravity-induced root bending is absent,apical hook formation still takes place.In such scenarios,hook formation begins with a de novo growth asymmetry at the apical part of a straightly elongating hypocotyl.Remarkably,suchde novo asymmetric growth,but not the following hook enlargement,precedes the establishment of a detectable auxin response asymmetry,and is largely independent of auxin biosynthesis,transport and signaling.Moreover,we found that functional cortical microtubule array is essential for the following enlargement of hook curvature.When microtubule array was disrupted by oryzalin,the polar localization of PIN proteins and the formation of an auxin maximum became impaired at the to-be-hook region.Taken together,we propose a more comprehensive model for apical hook initiation,in which the microtubuledependent polar localization of PINs may mediate the instruction of growth asymmetry that is either stochastically taking place,induced by gravitropic response,or both,to generate a significant auxin gradient that drives the full development of the apical hook.展开更多
Apical hook is a simple curved structure formed at the upper part of hypocotyls when dicot seeds germinate in darkness.The hook structure is transient but essential for seedlings’survival during soil emergence due to...Apical hook is a simple curved structure formed at the upper part of hypocotyls when dicot seeds germinate in darkness.The hook structure is transient but essential for seedlings’survival during soil emergence due to its efficient protection of the delicate shoot apex from mechanical injury.As a superb model system for studying plant differential growth,apical hook has fascinated botanists as early as the Darwin age,and significant advances have been achieved at both the morphological and molecular levels to understand how apical hook development is regulated.Here,we will mainly summarize the research progress at these two levels.We will also briefly compare the growth dynamics between apical hook and hypocotyl gravitropic bending at early seed germination phase,with the aim to deduce a certain consensus on their connections.Finally,we will outline the remaining questions and future research perspectives for apical hook development.展开更多
Spermatozoa emerging from the testis undergo a maturation process in the epididymis during which they change morphologically, biochemically and physiologically to gain motility and the ability to fertilize ova. We exa...Spermatozoa emerging from the testis undergo a maturation process in the epididymis during which they change morphologically, biochemically and physiologically to gain motility and the ability to fertilize ova. We examined mouse epididymal sperm with immunostaining and transmission electron microscopy （EM） and identified a previously unknown structure on the apical hook. The structure has a coiled configuration around 11 nm in thickness and is present at the tip of each corner of the triangular-rod shaped perforatorium. Surveying sperm isolated from various regions of the epididymis indicated that mouse sperm acquire the hook rim （HR） structure during its passage through the proximal two-thirds of the caput epididymidis. The structure withstands vigorous sonication and harsh chemical treatments and remains intact after the acrosome reaction. Its location and sturdiness suggest a function in protecting the apical hook from mechanical wear during fertilization. Our EM images of epididymal sperm also revealed additional novel structures as well as lateral asymmetry of the sperm head, indicating that mouse sperm head has a structure more complex than previously recognized.展开更多
文摘The Arabidopsis XBAT35 is one of five structurally related ankyrin repeat-containing Really Interesting New Gene （RING） E3 ligases involved in ubiquitin-mediated protein degradation, which plays key roles in a wide range of cel- lular processes. Here, we show that the XBAT35 gene undergoes alternative splicing, generating two transcripts that are constitutively expressed in all plant tissues. The two splice variants derive from an exon skipping event that excludes an in-frame segment from the XBAT35 precursor mRNA, giving rise to two protein isoforms that differ solely in the presence of a nuclear localization signal （NLS）. Transient expression assays indicate that the isoform lacking the NLS localizes in the cytoplasm of plant cells, whereas the other is targeted to the nucleus, accumulating in nuclear speckles. Both isoforms are functional E3 ligases, as assessed by in vitro ubiquitination assays. Two insertion mutant alleles and RNA-interference （RNAi） silencing lines for XBAT35 display no evident phenotypes under normal growth conditions, but exhibit hyper- sensitivity to the ethylene precursor 1-aminocyclopropane-l-carboxylate （ACC） during apical hook exaggeration in the dark, which is rescued by an inhibitor of ethylene perception. Independent expression of each XBAT35 splice variant in the mutant background indicates that the two isoforms may differentially contribute to apical hook formation but are both functional in this ethylene-mediated response. Thus, XBAT35 defines a novel player in ethylene signaling involved in negatively regulating apical hook curvature, with alternative splicing controlling dual targeting of this E3 ubiquitin ligase to the nuclear and cytoplasmic compartments.
基金funded by the Southern University of Science and Technology for scientific research start-ups(Grant No.Y01226124 to H.G.)National Natural Science Foundation of China(Grant No.31700239 to Y.W.)+1 种基金Shenzhen Science and Technology Innovation Program(Grant No.JCYJ20170817105503416 to W.L.)Key Laboratory of Molecular Design for Plant Cell Factory of Guangdong Higher Education Institutes(SUSTech)(2019KSYS006 to H.G.)。
文摘The development of a hook-like structure at the apical part of the soil-emerging organs has fascinated botanists for centuries,but how it is initiated remains unclear.Here,we demonstrate with highthroughput infrared imaging and 2-D clinostat treatment that,when gravity-induced root bending is absent,apical hook formation still takes place.In such scenarios,hook formation begins with a de novo growth asymmetry at the apical part of a straightly elongating hypocotyl.Remarkably,suchde novo asymmetric growth,but not the following hook enlargement,precedes the establishment of a detectable auxin response asymmetry,and is largely independent of auxin biosynthesis,transport and signaling.Moreover,we found that functional cortical microtubule array is essential for the following enlargement of hook curvature.When microtubule array was disrupted by oryzalin,the polar localization of PIN proteins and the formation of an auxin maximum became impaired at the to-be-hook region.Taken together,we propose a more comprehensive model for apical hook initiation,in which the microtubuledependent polar localization of PINs may mediate the instruction of growth asymmetry that is either stochastically taking place,induced by gravitropic response,or both,to generate a significant auxin gradient that drives the full development of the apical hook.
基金supported by National Natural Science Foundation of China(32230008 to H.G.)Shenzhen Science and Technology Program(KQTD20190929173906742 to H.G.)Key Laboratory of Molecular Design for Plant Cell Factory of Guangdong Higher Education Institutes(2019KSYS006 to H.G.)。
文摘Apical hook is a simple curved structure formed at the upper part of hypocotyls when dicot seeds germinate in darkness.The hook structure is transient but essential for seedlings’survival during soil emergence due to its efficient protection of the delicate shoot apex from mechanical injury.As a superb model system for studying plant differential growth,apical hook has fascinated botanists as early as the Darwin age,and significant advances have been achieved at both the morphological and molecular levels to understand how apical hook development is regulated.Here,we will mainly summarize the research progress at these two levels.We will also briefly compare the growth dynamics between apical hook and hypocotyl gravitropic bending at early seed germination phase,with the aim to deduce a certain consensus on their connections.Finally,we will outline the remaining questions and future research perspectives for apical hook development.
文摘Spermatozoa emerging from the testis undergo a maturation process in the epididymis during which they change morphologically, biochemically and physiologically to gain motility and the ability to fertilize ova. We examined mouse epididymal sperm with immunostaining and transmission electron microscopy （EM） and identified a previously unknown structure on the apical hook. The structure has a coiled configuration around 11 nm in thickness and is present at the tip of each corner of the triangular-rod shaped perforatorium. Surveying sperm isolated from various regions of the epididymis indicated that mouse sperm acquire the hook rim （HR） structure during its passage through the proximal two-thirds of the caput epididymidis. The structure withstands vigorous sonication and harsh chemical treatments and remains intact after the acrosome reaction. Its location and sturdiness suggest a function in protecting the apical hook from mechanical wear during fertilization. Our EM images of epididymal sperm also revealed additional novel structures as well as lateral asymmetry of the sperm head, indicating that mouse sperm head has a structure more complex than previously recognized.
