cDNA cloning, nucleotide sequence and expression of the gene for arylsulfatase in the sea urchin (Hemicentrotus pulcherrimus) embryo

Sasaki, Hiroshi; Yamada, Kazumi; Akasaka, Koji; Kawasaki, Hiroshi; Suzuki, Koichi; Saito, Akiko; Sato, Moriyuki; Shimada, Hiraku

doi:10.1111/j.1432-1033.1988.tb14338.x

Cited by 64 publications

(23 citation statements)

References 24 publications

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“…O/A patterning has been studied most in the ectoderm and, thus, many gene products with asymmetric distribution about the axis have been identified. Aboral genes include Spec1 and Spec2 (Lynn et al, 1983), CyIIIa actin (Cox et al, 1986), arylsulfatase (Sasaki et al, 1988), Hox8/Hbox1 (Angerer et al, 1989), SpMTA (Nemer et al, 1995) and P3A2 (Calzone et al, 1991); whereas oral genes include EctoV (Coffman and McClay, 1990) Otx (Li et al, 1997a;Yuh et al, 2002), SpCOUP-TF (Vlahou et al, 1996), BMP2/4 , PlOtp (Di Bernardo et al, 1999), Brachyury (Gross and McClay, 2001) and goosecoid (Angerer et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

LvTbx2/3: a T-box family transcription factor involved in formation of the oral/aboral axis of the sea urchin embryo

Gross

Peterson

et al. 2003

Development

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T-box family transcription factors have been identified in many organisms and are frequently associated with patterning events during embryonic development. With an interest in the molecular basis of patterning in the sea urchin embryo, we identified several members of the T-box family inLytechinus variegatus. Here, we report the cloning and characterization of an ortholog of the Tbx2/3 subfamily, LvTbx2/3. To characterize the spatial distribution of LvTbx2/3 protein throughout sea urchin embryogenesis, a polyclonal antiserum was generated. Nuclear localization of LvTbx2/3 initiated at the mesenchyme blastula stage and protein was present into the pluteus stage. Localization was asymmetric throughout this period and costaining with marker genes indicated that asymmetry was about the oral/aboral (O/A) axis. Asymmetric distribution of LvTbx2/3 was observed in the aboral territories of all three germ layers. In the skeletogenic mesoderm lineage, LvTbx2/3 expression was dynamic because expression appeared initially in all skeletogenic mesenchyme cells (PMCs) but,subsequently, became refined solely to the aboral ones during skeletogenesis. To determine if the aboral expression of LvTbx2/3 is linked between germ layers, and to place LvTbx2/3 in the sequence of events that specifies the O/A axis, the effects of a series of perturbations to O/A polarity on LvTbx2/3 expression in each germ layer were examined. Preventing the nuclear localization of β-catenin, pharmacological disruption of the O/A axis with NiCl2, overexpression ofBMP2/4 and disruption of the extracellular matrix all blockedLvTbx2/3 expression in all germ layers. This indicates that expression of LvTbx2/3 in the aboral territories of each germ layer is a common aspect of O/A specification, downstream of the molecular events that specify the axis. Furthermore, blocking the nuclear localization ofβ-catenin, overexpression of BMP2/4 and disruption of the extracellular matrix also prevented the oral (stomodael) expression of LvBrachyury (LvBrac) protein, indicating that the O/A axis is established by a complex series of events. Last, the function of LvTbx2/3 in the formation of the O/A axis was characterized by examining the phenotypic consequences of ectopic expression of LvTbx2/3 mRNA on embryonic development and the expression of marker genes that identify specific germ layers and tissues. Ectopic expression of LvTbx2/3 produced profound morphogenetic defects in derivatives of each germ layer with no apparent loss in specification events in those tissues. This indicates that LvTbx2/3 functions as a regulator of morphogenetic movements in the aboral compartments of the ectoderm, endoderm and mesoderm.

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Section: Introductionmentioning

confidence: 99%

LvTbx2/3: a T-box family transcription factor involved in formation of the oral/aboral axis of the sea urchin embryo

Gross

Peterson

et al. 2003

Development

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“…In heterokaryon studies of an electrophoretic variant of arylsulfatase, it was shown that the cys-3+ gene product could activate arylsulfatase gene expression in a trans-nuclear manner, whereas scon+ was limited to intranuclear control of arylsulfatase expression (6,26,28). Recent molecular studies with a number of arylsulfatases (e.g., human [45] and sera urchin [40] …”

mentioning

confidence: 99%

Molecular cloning and regulatory analysis of the arylsulfatase structural gene of Neurospora crassa.

Paietta

1989

Mol. Cell. Biol.

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The ars-l+ gene of Neurospora crassa encodes the enzyme arylsulfatase. ars-l+ is in a group of highly regulated sulfur-related structural genes that are expressed under conditions of sulfur limitation and are under coordinate control of the cys-3+ and scon+ regulatory genes. The ars-l+ gene was cloned by chromosome walking from the qa gene cluster, using a lambda library. Cotransformation of an N. crassa ars-1 mutant with the isolated lambda clones and the benomyl resistance gene, followed by assay for arylsulfatase activity, was used to screen for the ars-l+ gene. Further confirmation that the cloned segment mapped to the ars-l+ locus was obtained by restriction-fragment-length polymorphism analysis. Northern (RNA) blot analysis showed that the ars-l + gene was transcribed to give an mRNA of 2.3 kilobases. In wild-type cells, the ars-1 + transcript was abundant under sulfur-derepressing conditions but absent under repressing conditions. Time course analysis showed that the appearance of ars-l+ message in sulfur-derepressed cultures paralleled the appearance of arylsulfatase enzyme activity. In addition, transcription of ars-l+ was detected only under derepressing conditions in a nuclear transcription assay. In a cys-3 regulatory mutant that was unable to synthesize arylsulfatase (or other sulfur-controlled enzymes), there was no ars-l+ transcript under repressing or derepressing conditions. In a temperature-sensitive cys-3 mutant, the ars-l + transcript was present only at the permissive growth temperature and under sulfur derepression. A negative regulatory mutant, sconC, displayed both constitutive expression of arylsulfatase enzyme activity and content of ars-l + message.In the filamentous fungus Neurospora crassa, the synthesis of sets of enzymes in global areas of metabolism such as nitrogen, phosphorus, and sulfur metabolism is highly regulated (24, 26). The sulfur regulatory circuit of N. crassa provides a model system for studying coordinate gene regulation and how a cell regulates its sulfur status. The sulfur control system is composed of a genetically defined set of trans-acting regulatory genes and a set of structural genes that encode enzymes used in the uptake and assimilation of a variety of sulfur compounds (6,26). When cultures of N. crassa are grown under conditions of sulfur limitation (i.e., derepressing conditions), then the set of sulfur-related genes is expressed in a coordinate manner. The structural genes involved encode for arylsulfatase, choline sulfatase, choline sulfate permease, methionine permease, sulfate permeases I and II, and an extracellular protease. None of the known genes in the sulfur regulatory circuit are linked (14,19,23,36).An important part of the sulfur control system involves the positive regulatory gene, cys-3+. In cys-3 mutants there is a pleiotropic loss of the entire set of sulfur-controlled enzymes (25). Paietta et al. (34) cloned the cys-3+ regulatory gene, and subsequent work (12) has shown that the encoded product is a 25.9-kilodalton protein that contains ...

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“…The Ars gene is actively transcribed during post gastrula stages, and the accumulation of arylsulfatase mRNA may reach about 1% of total mRNAs (Sasaki et al, 1988); the C15 enhancer in the first intron is apparently very strong. Therefore, the Ars enhancer could influence the promoter of the neighboring genes.…”

Section: Boundary Of Chromatinmentioning

confidence: 99%

“…HpArs (Sakamoto et al, 1997;Kiyama et al, 1998), Spec2a (Gan et al, 1995) and Endo16 (Yuh et al, 1998) have been reported as direct target genes of Otx in sea urchin embryo. HpArs is transcriptionally activated late in blastula, and after the gastrula stage it is expressed exclusively in the aboral ectoderm throughout embryonic development (Sasaki et al, 1988;Akasaka et al, 1990aAkasaka et al, , 1990b. In order to detect cis-regulatory elements responsible for the transcriptional activity of genes, we introduce reporter(Luc)-fusion constructs into fertilized eggs.…”

Section: Otx Target Genementioning

confidence: 99%

“…We have been studying the mechanism of expression of Ars gene, which is expressed in a tightly regulated manner (Sasaki et al 1987, Sasaki et al, 1988Akasaka et al, 1990a, b) during sea urchin development. The Ars gene is actively transcribed during post gastrula stages, and the accumulation of arylsulfatase mRNA may reach about 1% of total mRNAs (Sasaki et al, 1988); the C15 enhancer in the first intron is apparently very strong.…”

Section: Boundary Of Chromatinmentioning

confidence: 99%

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A Sea Cucumber Homolog of the MouseT-Brain-1is Expressed in the Invaginated Cells of the Early Gastrula in Holothuria leucospilota

Maruyama¹

2000

Zoological Science

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cDNA cloning, nucleotide sequence and expression of the gene for arylsulfatase in the sea urchin (Hemicentrotus pulcherrimus) embryo

Cited by 64 publications

References 24 publications

LvTbx2/3: a T-box family transcription factor involved in formation of the oral/aboral axis of the sea urchin embryo

LvTbx2/3: a T-box family transcription factor involved in formation of the oral/aboral axis of the sea urchin embryo

Molecular cloning and regulatory analysis of the arylsulfatase structural gene of Neurospora crassa.

A Sea Cucumber Homolog of the MouseT-Brain-1is Expressed in the Invaginated Cells of the Early Gastrula in Holothuria leucospilota

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