The Molecular Biology of Artemia Haemoglobins

Manning, Anthony M.; Marshall, Craig J.; Powell, Robert; Trotman, C. N. A.; Tate, Warren P.

doi:10.1007/978-1-4757-0004-6_44

Cited by 2 publications

(2 citation statements)

References 10 publications

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“…However, this behaviour does not reflect a more general resumption of transcriptional activity in the developing cyst, since equivalent amounts of total RNA and poly(A)-containing mRNA have been obtained from dormant and 12 h-developed Artemia embryos (Wahba & Woodley, 1984). Moreover, the expression of histone (Bagshaw, 1982) and globin (Manning et al, 1989) genes takes place only in the nauplius stage, after 24 h of development. Thus protein synthesis should play a major role in executing the early developmental programme, since any changes in biochemical parameters reflect differentiation of preexisting cells (Hentschel & Tata, 1976).…”

Section: Introductionmentioning

confidence: 97%

Characterization of cell-free protein-synthesis systems from undeveloped and developing Artemia embryos

Moreno

Méndez

Haro

1991

Biochemical Journal

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We have developed and characterized translationally active cell-free systems from Artemia embryos at different developmental times. The optimized lysates from 16 h-developed embryos incorporated radiolabelled amino acids into polypeptides for up to 120 min. The polypeptides synthesized ranged in Mr from 150,000 to 10,000, suggesting that the endogenous mRNA was capable of directing the synthesis of complete polypeptides. Similar results were obtained by using lysates from early developmental stages; even the cell-free system prepared from 1 h-developed embryos was partially active in protein synthesis. Furthermore, all these lysates were capable of re-initiation, as demonstrated by inhibition of initiation with the inhibitors edeine and 7-methylguanosine 5'-triphosphate. Because we found no endogenous protein-synthetic activity in the corresponding lysates from undeveloped embryos, we have used cell-free translation systems from 0 h- and 16 h-developed Artemia embryos to analyse the mechanisms limiting protein synthesis at very early developmental stages. Undeveloped-embryo lysates supplemented with nuclease-treated reticulocyte lysate were capable of translating endogenous mRNAs to give products with a wide spectrum of Mr values, but lysates of 16 h-developed embryos supplemented in this way were not further stimulated. The nuclease-treated lysate appeared to be unnecessary 5 h after resumption of development. These results suggested that a component(s) limiting translation in the undeveloped-embryo lysate was provided by the nuclease-treated reticulocyte lysate, and that this component(s) no longer limited protein synthesis after development. In view of these results, partially fractionated reticulocyte lysates were tested for restoration of protein-synthetic activity in the undeveloped embryo lysate. A high-salt ribosomal wash devoid of ribosomal subunits, which is considered a crude polypeptide-initiation-factor preparation, also restored translation activity in the undeveloped embryo lysate and made it capable of directing the synthesis of both endogenous mRNAs and exogenous (globin) mRNA.

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Section: Introductionmentioning

confidence: 97%

Characterization of cell-free protein-synthesis systems from undeveloped and developing Artemia embryos

Moreno

Méndez

Haro

1991

Biochemical Journal

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“…Artemia mRNA was extracted from 44-h-old nauplii and cDNA was cloned and sequenced as previously described (Manning et al 1989(Manning et al , 1990a.…”

Section: Methodsmentioning

confidence: 99%

Interdomain linkage in the polymeric hemoglobin molecule of Artemia

et al. 1994

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Artemia has evolved the longest known concatenation of hemoglobin domains, the alpha subunit containing nine domains and the beta subunit having a similar size. Translation of the cDNA sequence of the alpha subunit reveals eight regions of inter-domain polypeptide linking together the nine heme-binding domains, together with partially analogous sequences preceding the first domain and following the last. Analysis of the structural possibilities of the linker sequences suggests how the domains may be organized in the subunit. The interdomain linker sequences were 14%-64% identical (62%-91% similar by Dayhoff substitution matrix) and approximately 14 residues in length including a consensus -Val-Asp-Pro-Val-Thr-Gly-Leu-. The linker composition resembled that of the 11 amino acid pre-A leader sequence of Petromyzon marinus (lamprey) hemoglobin V, the structure of which is known. Prediction of structure from the Artemia linker sequences indicated a nonhelical, turn-associated linker which could be modeled to the Petromyzon leader. Measurements confirmed that such a structure could support the packing of nine Artemia domains into a polymeric subunit of annular shape, two of which subunits (which can be similar or dissimilar) comprise the physiological molecule.(ABSTRACT TRUNCATED AT 250 WORDS)

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The Molecular Biology of Artemia Haemoglobins

Cited by 2 publications

References 10 publications

Characterization of cell-free protein-synthesis systems from undeveloped and developing Artemia embryos

Characterization of cell-free protein-synthesis systems from undeveloped and developing Artemia embryos

Interdomain linkage in the polymeric hemoglobin molecule of Artemia

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