Accumulation and decay of DG42 gene products follow a gradient pattern during Xenopus embryogenesis

Rosa, Frédéric; Sargent, Thomas D.; Rebbert, Martha L.; Michaels, George S.; Jamrich, Milan; Grunz, Horst; Jónás, Erzsébet; Winkles, Jeffrey A.; Dawid, Igor B.

doi:10.1016/0012-1606(88)90166-2

Cited by 98 publications

(68 citation statements)

References 36 publications

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“…Briefly, PCR utilizing Vent DNA polymerase (New England Biolabs) was used to create separate xlHAS1 (from the cDNA plasmid template, pC4202; Ref. 7) or GFP (from the pGLO plasmid template; Bio-Rad) amplicons. The primer pairs were HAS-sense, (5Ј-GGAATTCATGAAAGAAAAAGCTGCAGA-AACCATGG-3Ј) and HAS-antisense (5Ј-ATGGTGATGGTGGTGGTG-3Ј) and GFP-sense (5Ј-ATGGCTAGCAAAGGAGAAGAAC-3Ј) and GFPantisense (5Ј-GCACGCGGCCGCTGCAGTTATTTGTAGAGCTCATCC-ATGC-3Ј) (Great American Gene Company).…”

Section: Production Of Recombinant Native and Fusion Xlhas1mentioning

confidence: 99%

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Functional Molecular Mass of a Vertebrate Hyaluronan Synthase as Determined by Radiation Inactivation Analysis

Pummill¹,

Kempner²,

DeAngelis³

2001

Journal of Biological Chemistry

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Hyaluronan (HA), a linear polysaccharide composed of N-acetylglucosamine-glucuronic acid repeats, is found in the extracellular matrix of vertebrate tissues as well as the capsule of several pathogenic bacteria. The HA synthases (HASs) are dual-action glycosyltransferases that catalyze the addition of two different sugars from UDP-linked precursors to the growing HA chain. The prototypical vertebrate hyaluronan synthase, xl-HAS1 (or DG42) from Xenopus laevis, is a 588-residue membrane protein. Recently, the streptococcal enzyme was found to function as a monomer of protein with ϳ16 lipid molecules. The vertebrate enzymes are larger than the streptococcal enzymes; based on the vertebrate HAS deduced amino acid sequence, two additional membrane-associated regions at the carboxyl terminus are predicted. We have utilized radiation inactivation to measure the target size of yeast-derived recombinant xlHAS1. The target size of HAS activity was confirmed using two internal standards. First, samples were spiked with glucose-6-phosphate dehydrogenase, an enzyme of known molecular weight. Second, parallel samples of native xlHAS1 and a xlHAS1-green fluorescent protein fusion (833 residues) were compared; substantial confidence was gained by using this novel internal standard. Our test also corroborated the basic tenets of radiation inactivation theory. We found that the vertebrate HAS protein functions catalytically as a monomer.

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Section: Production Of Recombinant Native and Fusion Xlhas1mentioning

confidence: 99%

“…A Xenopus laevis (African clawed frog) protein, xlHAS1, originally called DG42 (for differentially expressed in gastrulation), was the first vertebrate HAS to be cloned, but at the time, its function was not known (7). The amino acid sequence of xlHAS1 is quite similar to that of bacterial spHAS enzyme (3,4).…”

mentioning

confidence: 99%

Functional Molecular Mass of a Vertebrate Hyaluronan Synthase as Determined by Radiation Inactivation Analysis

Pummill¹,

Kempner²,

DeAngelis³

2001

Journal of Biological Chemistry

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show abstract

“…A high level of similarity has also been detected with the DG42 protein, which is transiently expressed during embryogenesis of Xenopus laevis (Rosa et al, 1988 ;Bulawa, 1992). NodC and DG42 function as N-acetylglucosamine transferases.…”

Section: mentioning

confidence: 91%

Expression of chitin synthase genes in the arbuscular mycorrhizal fungus Gigaspora margarita

Lanfranco¹,

Vallino²,

Bonfante³

1999

New Phytologist

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Genomic DNA from spores of the endomycorrhizal fungus Gigaspora margarita was amplified with two sets of degenerate primers designed on conserved regions of chitin synthase polypeptides. Three chitin synthase (chs) gene portions were identified : Gimchs1, Gimchs2 and Gimchs3, encoding one class II and two class IV chitin synthases, respectively. Gimchs1 and Gimchs3 transcripts were detected by reverse transcription polymerase chain reaction (RT-PCR) during root colonization, whereas none of three mRNAs was revealed during spore germination. The potential role of the genes expressed is discussed in the light of morphogenetic and signalling events.

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“…The prenylated vesicle-associated SNARE (v-SNARE) is localized to the ER-Golgi network and plays a role in membrane formation (Park et al, 1998). Xenopus DG42 is a ventrally expressed hyaluronan synthetase involved in the synthesis of glycosaminoglycans, which provide a hydrophilic medium in the extracellular space (Rosa et al, 1988;Wessely et al, 2004).…”

Section: Known Lysosomal/er/golgi Proteinsmentioning

confidence: 99%

Exploration of the extracellular space by a large-scale secretion screen in the early Xenopus embryo

et al. 2005

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Secreted proteins play a crucial role in intercellular communication during embryogenesis and in the adult. We recently described a novel method, designated as secretion cloning, that allows identifying extracellular proteins exclusively based on their ability to be secreted by transfected cells. In this paper, we present the results of a large-scale screening of more than 90,000 clones from three cDNA expression libraries constructed from early Xenopus embryos. Of 170 sequenced clones, 65 appeared to encode secreted proteins; 26 clones (40%) were identical to previously known Xenopus genes, 25 clones (38%) were homologous to other genes identified in various organisms and 14 clones (22%) were novel. Apart from these bona fide secreted proteins, we also isolated lysosomal or other secretory pathway proteins and some cytoplasmic proteins commonly found in body fluids. Among the novel secreted proteins were two putative growth factors of the Granulin family, termed xGra1 and xGra2; they are structurally similar to EGF and TGFα and show a spotted expression pattern in the epidermis. Another secreted protein, designated xSOUL, belongs to the family of heme-binding proteins and exhibits distinct expression in the early brain. A third protein, termed Xystatin, is related to cysteine proteinase inhibitors. Our results indicate that secretion cloning is an effective and generally useful tool for the unbiased isolation of secreted proteins.

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Accumulation and decay of DG42 gene products follow a gradient pattern during Xenopus embryogenesis

Cited by 98 publications

References 36 publications

Functional Molecular Mass of a Vertebrate Hyaluronan Synthase as Determined by Radiation Inactivation Analysis

Functional Molecular Mass of a Vertebrate Hyaluronan Synthase as Determined by Radiation Inactivation Analysis

Expression of chitin synthase genes in the arbuscular mycorrhizal fungus Gigaspora margarita

Exploration of the extracellular space by a large-scale secretion screen in the early Xenopus embryo

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