Translation of the messenger RNA for rabbit uteroglobin in <i>Xenopus</i> oocytes

Beato, Miguel

doi:10.1016/0014-5793(75)80398-x

Cited by 33 publications

(15 citation statements)

References 9 publications

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“…Ribosomal ribonucleic acids, 5s, 16s and 23s, from Escherichia coli, ribonuclease A from bovine pancreas, ribonuclease Ti from Aspergillus oryzae, proteinase K, adenylyl-(3t 5I) 5-adenosine, and adenylyl-(3'-.5')9-adenosine were obtained from Boehringer, Mannheim. t-RNA was prepared from rabbit liver or rabbit reticulocytes by the procedure of Rogg et al (4).…”

Section: Methodsmentioning

confidence: 99%

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Purification and properties of rabbit uterus preuteroglobin mRNA

1977

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The mRNA for preuteroglobin, a precursor of the hormonally induced protein uteroglobin, has been partially purified from the endometrium of progesterone treated rabbits. The purification procedure starts with total endometrial polysomes and involves treatment with proteinase K and dodecylsulfate, chromatography on oligo(dT)-cellulose, sucrose density gradient centrifugation, electrophoresis in polyacrylamide gels containing dodecylsulfate, and a second absorption to oligo(dT)-cellulose. The final mRNA preparation codes exclusively for preuteroglobin in a wheat germ cell-free system and migrates as a single band in polyacrylamide gels containing 99% formamide. The average length of the poly(A) segment is 60 nucleotides and the translation of the preuteroglobin mRNA is inhibited by m7G(5')ppp(5')A, indicating that it contains a "tcapped" 5'-terminus. Comparison with known standards yields a molecular weight of 200,000 (600 nucleotides) for preuteroglobin mRNA, approximately twice as many nucleotides as required for encoding the 90 aminoacids of its cell-free product.

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Section: Methodsmentioning

confidence: 99%

“…The RNA was then precipitated overnight at -200 after the addition of 0.1 volume of 2M LiCl and 2 volumes of ethanol. The RNA contained in the material which did not bind to oligo(dT)-cellulose was extracted with phenol/chloroform (5) and precipitated in a similar way.…”

Section: Methodsmentioning

confidence: 99%

Purification and properties of rabbit uterus preuteroglobin mRNA

1977

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“…In experiments using purified mRNAs (11,12,34,35) (6,37,38), whereas T antigens are synthesized early in infection and are required for the induction of host and viral DNA replication (39,40,41). Further experiments are needed to determine whether in the oocytes the control of late viral gene expression (i.e., synthesis of viral capsids) is similar to or different from that operating in mammalian cells undergoing lytic infection.…”

Section: Resultsmentioning

confidence: 99%

“…The procedure used for injecting viral DNA into the germinal vesicle of Xenopus oocytes was a modification of already established microinjection techniques for DNA (4, 10) and mRNA (11,12 24 hr (1 Ci = 3.7 X 1010 becquerels). The surviving oocytes (60-90%) were washed first with Barth's solution, then with distilled water, and stored at -80'C after removal of the water.…”

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confidence: 99%

DNAs of simian virus 40 and polyoma direct the synthesis of viral tumor antigens and capsid proteins in Xenopus oocytes.

Rungger¹,

Türler²

1978

Proc. Natl. Acad. Sci. U.S.A.

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Purified simian virus 40 and polyoma DNAs injected into nuclei of Xenopus Nuclei from various eukaryotic cells injected into frog oocyte nuclei (germinal vesicles) continue to direct synthesis of origin-specific proteins (1). In addition, genes that were not expressed in the donor cell may be activated after injection (2). Purified viral, phage, or plasmid DNAs injected into the nucleus of Xenopus oocytes serve as templates for the transcription of specific RNA sequences (3). The synthesis of 5S RNA (4) and tRNA (5) is greatly stimulated upon injection of the corresponding templates. It has been claimed that, after injection of simian virus 40 (SV40) DNA I (6), much of the virus-specific RNA synthesized in the Xenopus oocyte has the same size and is transcribed from the same region as the late viral mRNAs present in lytically infected monkey cells (7,8), and proteins that migrate on two-dimensional gels in the same manner as the SV40 capsid proteins VP 1 and VP 3 were detected (8).In this article we report that Xenopus oocytes transcribe and translate most or all of the genetic information contained in the SV40 genome. In particular, we observed the synthesis of large (T) and small (t) SV40 tumor antigens having the same characteristics as those synthesized in monkey cells undergoing lytic infection. Injection of polyoma DNA I (6) resulted in the synthesis of three polyoma-specific polypeptides comigrating, during gel electrophoresis, with two minor polypeptides related to polyoma tumor antigen and with the major capsid protein, respectively.MATERIALS AND METHODS SV40 DNA I (covalently closed circular) was isolated from SV40-infected CV-1 African green monkey cells (Flow Laboratories, Irvine, Scotland) 40-48 hr after infection by selective extraction, deproteinized with phenol, and purified by CsCl/ethidium bromide density gradient equilibrium centrifugation (9). Polyoma DNA I was isolated from polyomainfected mouse kidney cell cultures 30-35 hr after infection by the same procedure. For injection, the viral DNA was dissolved in 88 mM NaCl/1 mM KCI/15 mM Tris-HCl, pH 7.6 at concentrations between 300 and 500 Ag/ml. 24 hr in Barth's solution containing 50 units per ml each of streptomycin, penicillin, and kanamycin. Thereafter 1 ml of Barth's solution containing 300 ,uCi of [35S]methionine (500-1000 Ci/mmol, the Radiochemical Centre, Amersham, England) was added and incubation was continued for another 24 hr (1 Ci = 3.7 X 1010 becquerels). The surviving oocytes (60-90%) were washed first with Barth's solution, then with distilled water, and stored at -80'C after removal of the water.For extraction of viral proteins, the oocytes were disrupted in a small Dounce homogenizer in 2 ml of a buffer containing detergent (0.1 M Tris-HCl, pH 9/0.1 M NaCl/5 mM KCI/1 mM CaCl2/0.5 mM MgCl2/0.7 mM Na2HPO4/0.5% Nonidet P-40). The homogenate was sonicated in a MSE ultrasonic disintegrator (Mk 1) for 1 min with an amplitude of 10 Atm and kept on ice for 20 min. After centrifugation at 15,000 X g for 30 min at 40C the s...

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“…Endometrial uteroglobin can be induced by progesterone ad¬ ministration, which leads to an accumulation of the mRNA for a precursor of the uteroglobin monomer in the endometrial polysomes (Beier, 1968;Beier, Petry & Kiihnel, 1970;Bullock & Willen, 1974;Mayol & Longenecker, 1974;Beato & Arnemann, 1975;Beato & Rungger, 1975;Bullock et al, 1976;Levey & Daniel, 1976). In the oviduct, the uteroglobinlike protein is induced by oestrogens (Kay & Feigelson, 1972;Goswami & Feigelson, 1974), and in the lung its presence appears to be independent of ovarian steroids (Noske & Feigelson, 1976).…”

Section: Introductionmentioning

confidence: 99%