Biopolym. Cell. 2006; 22(3):192-200.
http://dx.doi.org/10.7124/bc.000730
Structure and Function of Biopolymers
Investigation of structure-function interaction of Bombyx mori nuclear polyhedrosis virus polyhedrin with polyhedra protease and RNA. Effect of protease on polyhedrin dissociation during Sepharose 6B gel-filtration
1Kozlov E. A., 1Nijazova N., 1Bobrovskaja M. T., 1Shirina T. V.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

It is known that B.mori NPV polyhedra contains protease which cleaves polyhedrin polypeptide chain (m.w. 28500) during dissolution of polyhedra in vitro under alkaline conditions (pH10.5) to overlapping fragments with mol. weights 25000, 22000, 19000, 10000 and 8000. Besides polyhedrin, present in solution at pH10.5 as high mol.weight 13S- (70 %) and >13S- (30 %) associates. Current paper shows that during Sepharose 6B gel-filtration at pH10.5 of polyhedrin pH 10,5 solution there occurs partial dissociation of high mol. weight associates to unknown earlier 8,5S-associates, 25000 polypeptide and its dimer with sedimentation coefficient 3,4S. Undissociated high mol. weight associates and 8.5S-associates contain the whole polypeptide chain (m.w. 28500) only. During gel-filtration at pH11.0 of polyhedrin solution with the same pH there occurs complete dissociation of high mol. weight associates to 8.5S-associates, 22000 polypeptide and its dimer with sedimentation coefficient 3.2S. The 8.5S-associate contains the whole 28500 polypeptide chain only. RNA was found in the low mol. weight (<25000) and high mol. weight (13S and higher) zones. 8.5S-associate does not contain RNA. After preliminary polyhedra heating at 80?–90?C the polyhedra protease does not cleave polyhedrin in pH10.5 solution and dissociation of high mol. weight associates to 8.5S-associates containing whole polypeptide chain occurs with much lower degree during gel-filtration at pH10.5. Yet as a result of gel-filtration RNP-complex is released with mol. weight of 20000–25000 and sedimentation coefficient 2.9S, containing polypeptide chain with mol. weight of 10000–12000. The effect of the polyhedra protease and RNA on in vivo polyhedrin dissociation and virus realization from polyhedra are discussed.
Keywords: nuclear polyhedrosis virus, polyhedrin, association-dissociation, polyhedra protease, polyhedra RNA, RNP-complex
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References

[1] Zhimerikin VN. Some properties of the virus strain Birdia diprionis "Tomsk". Insect viruses Eds. NN Vorob'ev, LM Tarasevich. M.: Nauka, 1975; 197 p.
[2] Kozlov EA, Soguliaeva VM, Levitina TL, Vereshchak V, Serebriany? SV. [Purification of the polyhedral protein of nuclear polyhedrosis virus of mulberry silkworm and association of the protein in solution]. Biokhimiia. 1969;34(4):679-85.
[3] Kozlov EA, Unguryanu N, Serebryanyy SB. The possible role of proteases in virions nuclear polyhedrosis virus of the silkworm. Viral diseases of plants and insects: Tr LSHA. Elgava, 1980; 121:45-8.
[4] Bobrovskaya MT, Kovalev VA, Shirina TV, Rudenko AV, Kozlov EA. Dynamics of cleavage of the Bombyx mori nuclear polyhedrosis virus polyhedrin by polyhedral protease and identification of cleaved bonds. Biopolym Cell. 2004; 20(3):207-14.
[5] Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969;244(16):4406-12.
[6] Bowen TJ. An introduction to ultracentrifugation. Wiley-Interscience, New York, 1971. 171 p.
[7] Maibaum WW. Determination of pentoses in nucleotides by the Bial reaction. Biokhimiia. 1945;10(4):353-9.
[8] Bergold GH. Die isolierung des poliedervirus und die natur der polieder. Z. Naturforsch. 1947; 2b:122-43.
[9] Kozlov EA, Levitina TL, Gusak NM. The primary structure of baculovirus inclusion body proteins. Evolution and structure-function aspects. Curr Top Microbiol Immunol. 1986;131:135-64.
[10] Harrap KA. The structure of nuclear polyhedrosis viruses. I. The inclusion body. Virology. 1972;50(1):114–23.
[11] Lylo VV, Serebryani SB. Hydrodynamic characteristics of the 12S-oligomer of the protein of inclusion bodies of a nuclear polyhedrosis virus of the silkworm (Bombyx mori). Biopolym Cell. 1988; 4(3):139-144, 150.
[12] Payne CC, Kalmakoff J. Alkaline protease associated with virus particles of a nuclear polyhedrosis virus: assay, purification, and properties. J Virol. 1978;26(1):84-92.
[13] Kozlov EA, Serebriany? SB. [Degradation of silkworm nuclear polyhedrosis virus]. Mikrobiol Zh. 1980;42(3):365-71.