Isolation of biologically active poly(A)+ and poly (A)- mRNA from germinating wheat embryos

Filimonov, N. G.; Agashkin, A. O.; Sablina, O. D.; Aitkhozhin, M. A.

doi:10.7124/bc.0001C8

Biopolym. Cell. 1986; 2(6):307-311.

http://dx.doi.org/10.7124/bc.0001C8

Structure and Function of Biopolymers

Isolation of biologically active poly(A)⁺ and poly (A)^- mRNA from germinating wheat embryos

1Filimonov N. G., 1Agashkin A. O., 1Sablina O. D., 1Aitkhozhin M. A.

Institute of Molecular Biology and Biochemistry, Academy of Sciences of the Kazakh SSR
Alma-Ata, USSR

Abstract

Biologically active poly (A)^- mRNA were isolated from polyribosomes and free cytoplasmic informosomes of germinating wheat embryos by benzoylated cellulose chromato-graphy. Purified fractions of poly (A) ^- mRNA of polyribosomes and free isoformosomes do not essentially differ from cytoplasmic poly (A) ⁺ mRNA by molecular-weight characteristics and have translational activity in the cell-free system of protein synthesis. It is shown that poly (A) ^-mRNA of polyribosomes and free informosomes equally stimulate incorporation of [³⁵S]methionine into acidinsoluble product, but the efficiency of their translation is much lower than that of poly (A)⁺ mRNA. The difference in the translation efficiency of cytoplasmic poly (A) ⁺ and poly (A) ^- mRNA of wheat embryos is obviously the metabolitic peculiarity of these mRNA and is not related to their isolation.

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References

[1] Ajtkhozhin MA, Akhanov AU, Doschanov KhI. Informosomes of germinating wheat embryos. FEBS Lett. 1973; 31(1):104-6.

[2] Ajtkhozhin MA, Akhanov AU. Release of mRNP-particles of the informosome type from polyribosomes of higher plant embryos. FEBS Lett. 1974;41(2):275-9.

[3] Spirin AS, Ajtkhozhin MA. Informosomes and polyribosome-associated proteins in eukaryotes. Trends Biochem Sci. 1985; 10(4):162-165.

[4] Filimonov NG, Aitkhozhin MA, Gazarian KG. Poly(A)-containing RNA from germinating wheat embryos. Mol Biol (Mosk). 1978;12(3):522-6.

[5] Filimonov NG, Martakova NA, Popov LS, Tarantul VZ, Aitkhozhin MA. Nucleotide sequence organization of nuclear DNA of wheat embryos. Biokhimiia. 1982;47(7):1198-207.

[6] Perry RP, La Torre J, Kelley DE, Greenberg JR. On the lability of poly(A) sequences during extraction of messenger RNA from polyribosomes. Biochim Biophys Acta. 1972;262(2):220-6.

[7] Maniatis T, Fritsch E, Sambrook J. Isolation, purification and analysis of mRNA from eukaryotic cells. Methods of genetic engineering. Moscow, Mir, 1984; 186-204.

[8] Gillam I, Millward S, Blew D, von Tigerstrom M, Wimmer E, Tener GM. The separation of soluble ribonucleic acids on benzoylated diethylaminoethylcellulose. Biochemistry. 1967;6(10):3043-56.

[9] Marcus A, Efron D, Weeks DP. The wheat embryo cell-free system. Methods Enzymol. 1974;30:749-54.

[10] Roberts WK. Use of benzoylated cellulose columns for the isolation of poly(adenylic acid) containing RNA and other polynucleotides with little secondary structure. Biochemistry. 1974;13(18):3677-82.

[11] Huez G, Marbaix G, Hubert E, Leclercq M, Nudel U, Soreq H, Salomon R, Lebleu B, Revel M, Littauer UZ. Role of the polyadenylate segment in the translation of globin messenger RNA in Xenopus oocytes. Proc Natl Acad Sci U S A. 1974;71(8):3143-6.