RNA synthesis in starfish embryos: developmental consequences of its inhibition by formycin

Isomura, Haruhiko; Itoh, Noriyuki; Ikegami, Susumu

doi:10.1016/0167-4781(89)90157-7

Cited by 9 publications

(13 citation statements)

References 14 publications

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“…We found that formycin, an analogue of adenosine, added to a culture of starfish embryos interfered with the formation of UTP and CTP, thereby inhibiting RNA synthesis and halting embryonic development at the early blastula stage. It was notable that the nucleus was present in each blastomere of the development-arrested embryo [6]. Similar results were obtained with albuside [4].…”

Section: Discussionsupporting

confidence: 84%

“…At given times, aliquots were removed and the embryos were washed with cold (4°C) ASW and fixed in cold 5% (w/v) trichloroacetic acid (TCA). Radioactivity in aliquots of acid-soluble fractions was measured as described previously [6]. Radioactivity incorporated into UTP was measured by counting the UTP fraction, which was separated from other acid-soluble fractions by HPLC as described [6].…”

Section: Incorporation Of [~H]uridine Into Utp and Rnamentioning

confidence: 99%

“…They were suspended in 3 ml of 0.5 N NaOH and digested at 37°C overnight. Then 0.3 ml of 100% (w/v) TCA was added and the acid-soluble radioactivity was counted as described previously [6]. From these data the amounts of UMP incorporated into RNA werc calculated.…”

Section: Incorporation Of [~H]uridine Into Utp and Rnamentioning

confidence: 99%

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Selective inhibition of gastrulation in the starfish embryo by albuside B, an inosine analogue

et al. 1995

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External application of 0.2-100 pglml albuside B inhibits gastrulation of the starfish (Asterina pectinifera) embryo.Treated embryos retain the late blastula morphology with the vegetal plate. However, the vegetal plate is unreactive to soybean agglutinin, a probe for observing the progenitor cells of the archenteron (mesendoderm) in a normal embryo. The effective period of the treatment is limited from 4 to 6 h after fertilization, a period immediately before the onset of blastulation. RNA synthesis is unaffected during the period of sensitivity. The selectivity of the inhibition shows that albuside B may be a useful tool for studying the mechanisms of mesendoderm differentiation.

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Section: Discussionsupporting

confidence: 84%

Section: Incorporation Of [~H]uridine Into Utp and Rnamentioning

confidence: 99%

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Selective inhibition of gastrulation in the starfish embryo by albuside B, an inosine analogue

et al. 1995

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“…Eggs and sperm were obtained as described previously [18][19][20]. Eggs were fertilized and embryos were cultured in artificial sea water that contained 5 mgAEmL )1 streptomycin sulfate and 50 lgAEmL )1 penicillin G. Cultures were maintained in jars at a density of < 5000 embryos per mL with gentle stirring.…”

Section: Cultivation Of Embryosmentioning

confidence: 99%

Molecular characterization of a novel nuclear transglutaminase that is expressed during starfish embryogenesis

Sugino¹,

Terakawa²,

Yamasaki³

et al. 2002

Eur J Biochem

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We report the constitution and molecular characterization of a novel transglutaminase (EC 2.3.2.13) that starts to accumulate specifically in the nucleus in the starfish (Asterina pectinifera) embryo after progression through the early blastula stage. The cDNA for the nuclear transglutaminase was cloned and the cDNA-deduced sequence defines a single open reading frame encoding a protein with 737 amino acids and a predicted molecular mass of 83 kDa. A comparison of this transglutaminase with other members of the gene family revealed an overall sequence identity of 33-41%. A special sequence feature of this transglutaminase, which is not found in other transglutaminases, is the presence of nuclear localization signal-like sequences in the N-terminal region.Microinjection of hybrid constructs that encode the N-terminal segment fused to reporter proteins into the germinal vesicle of an oocyte produced chimeric proteins by transcription-coupled translation. It was found that the N-terminal segment alone was sufficient to effect nuclear accumulation of an otherwise cytoplasmic protein. These results suggest that the nuclear accumulation of the transglutaminase may play an important role in nuclear remodeling during early starfish embryogenesis.Keywords: transglutaminase; nucleus; starfish; embryo; cloning.The class of enzymes that are commonly referred to as transglutaminases (TG) (EC 2.3. Recent findings have shown that, apart from their protein modifying capabilities, tissue TG is also able to function as a component of the signal-transducing G protein complex [7]. The cDNA of G ha , involved in the transmission of adrenergic stimuli, is identical to that of tissue TG of human endothelial cells [7]. Tissue TG is localized mainly in the cytosol, but detectable tissue TG expression has been reported in the nucleus [8][9][10]. However, TG activity in the nucleus and the mechanisms of its translocation is not well understood, and nucleus-specific TG has not been reported.It is accepted that many proteins are able to cross nuclear membranes and accumulate against gradients to concentrate in the nucleus [11,12]. The nuclear translocation of proteins via the nuclear pore complex is dependent on a nuclear localization signal in the protein, which is rich in basic amino acids and may be bipartite [13][14][15]. The functional assays of such nuclear localization signals are usually based on the ability of a signal to confer nuclear localization to an otherwise non-nuclear protein.The present paper describes the occurrence of a novel TG that is localized exclusively in the nucleus of starfish (Asterina pectinifera) embryonic cells and is designated nuclear TG (nTG). The amino-acid sequence derived from the cDNA sequence contains putative nuclear localization signals [15] in the N-terminal region. We demonstrate here that the N-terminal region promotes the nuclear accumulation of an otherwise cytoplasmic protein, namely pyruvate kinase (PK), in the A. pectinifera oocyte system. This finding suggests that nuclear localization sig...

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“…The inhibitory activity of the sponge extract and various fractions purified from it was measured by adding a small number of fertilized eggs of A. pectinifera to serially diluted sample solutions and observing the cytological changes in the embryos cultured at 20°C as described. 6 ) One unit of activity was defined as the maximum volume (in ml) of a solution which showed sufficient effectiveness in arresting development at the preblastulation stage.…”

Section: Methodsmentioning

confidence: 99%

Effects of Okadaic Acid on Embryonic Development of the StarfishAsterina pectinifera

Ikegami

Kajiyama

Ozaki

et al. 1992

Bioscience, Biotechnology, and Biochemistry

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RNA synthesis in starfish embryos: developmental consequences of its inhibition by formycin

Cited by 9 publications

References 14 publications

Selective inhibition of gastrulation in the starfish embryo by albuside B, an inosine analogue

Selective inhibition of gastrulation in the starfish embryo by albuside B, an inosine analogue

Molecular characterization of a novel nuclear transglutaminase that is expressed during starfish embryogenesis

Effects of Okadaic Acid on Embryonic Development of the StarfishAsterina pectinifera

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