Isolation and Characterization of an Extremely Basic Protein from Adenovirus Type 5

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 1976 Mar 1

PMID 1255865

Citations 19

Authors

K Hosokawa

M T Sung

Affiliations

Soon will be listed here.

Abstract

By starch-gel electrophoresis and a staining method that is highly sensitive for argininyl residues, adenovirus type 5 was found to contain two minor basic polypeptides of extreme cathodic mobility in addition to the two known core proteins. The fastest-migrating polypeptide, named mu protein, and the second fastest polypeptide are found in adenovirions and virus-infected KB cells but not in top components or in uninfected cells. The top components and infected cells contain an additional basic polypeptide, presumably P-VII, that migrates slightly slower than polypeptide VII. None of the basic polypeptides of adenovirions was electrophoretically identical to the host histone. The basic proteins of adenovirions were purified by urea phosphocellulose column chromatography and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The two minor basic core proteins, mu and another component, have similar mobilities in sodium dodecyl sulfate-polyacrylamide gels as a complex of polypeptides X-XII. After further purification on a Sephadex G-75 column, the mu protein was found to have a molecular weight of about 4,000. Amino acid analysis showed that the mu protein lacks tryptophan and 69% of the total amino acid residues are basic, that is, 54% arginine, 13% histidine, and 2% lysine. Only eight amino acids seem to contribute to make the mu polypeptide. There are 125 copies of the mu polypeptide per 1,000 copies of polypeptide VII in a virion.

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References

CRAWFORD L . A study of the Rous sarcoma virus by density gradient centrifugation. Virology. 1960; 12:143-53. DOI: 10.1016/0042-6822(60)90190-2. View

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

Lewis J, Atkins J, Anderson C, Baum P, GESTELAND R . Mapping of late adenovirus genes by cell-free translation of RNA selected by hybridization to specific DNA fragments. Proc Natl Acad Sci U S A. 1975; 72(4):1344-8. PMC: 432530. DOI: 10.1073/pnas.72.4.1344. View

Brown D, Westphal M, Burlingham B, Winterhoff U, DOERFLER W . Structure and composition of the adenovirus type 2 core. J Virol. 1975; 16(2):366-87. PMC: 354676. DOI: 10.1128/JVI.16.2.366-387.1975. View

Prage L, Pettersson U, Hoglund S, LONBERG-HOLM K, Philipson L . Structural proteins of adenoviruses. IV. Sequential degradation of the adenovirus type 2 virion. Virology. 1970; 42(2):341-58. DOI: 10.1016/0042-6822(70)90278-3. View

Everitt E, Sundquist B, Pettersson U, Philipson L . Structural proteins of adenoviruses. X. Isolation and topography of low molecular weight antigens from the virion of adenovirus type 2. Virology. 1973; 52(1):130-47. DOI: 10.1016/0042-6822(73)90404-2. View

Laver W . Isolation of an arginine-rich protein from particles of adenovirus type 2. Virology. 1970; 41(3):488-500. DOI: 10.1016/0042-6822(70)90170-4. View

Rouse H, SCHLESINGER R . An arginine-dependent step in the maturation of type 2 adenovirus. Virology. 1967; 33(3):513-22. DOI: 10.1016/0042-6822(67)90128-6. View

Rouse H, SCHLESINGER R . The effects of arginine starvation on macromolecular synthesis in infection with type 2 adenovirus. I. Synthesis and utilization of structural proteins. Virology. 1972; 48(2):463-71. DOI: 10.1016/0042-6822(72)90057-8. View

10.

BABLANIAN R, Russell W . Adenovirus polypeptide synthesis in the presence of non-replicating poliovirus. J Gen Virol. 1974; 24(2):261-79. DOI: 10.1099/0022-1317-24-2-261. View

11.

Anderson C, Lewis J, Atkins J, GESTELAND R . Cell-free synthesis of adenovirus 2 proteins programmed by fractionated messenger RNA: a comparison of polypeptide products and messenger RNA lengths. Proc Natl Acad Sci U S A. 1974; 71(7):2756-60. PMC: 388549. DOI: 10.1073/pnas.71.7.2756. View

12.

Russell W, Skehel J . The polypeptides of adenovirus-infected cells. J Gen Virol. 1972; 15(1):45-57. DOI: 10.1099/0022-1317-15-1-45. View

13.

Walter G, Maizel Jr J . The polypeptides of adenovirus. IV. Detection of early and late virus-induced polypeptides and their distribution in subcellular fractions. Virology. 1974; 57(2):402-8. DOI: 10.1016/0042-6822(74)90180-9. View

14.

Ishibashi M, Maizel Jr J . The polypeptides of adenovirus. V. Young virions, structural intermediate between top components and aged virions. Virology. 1974; 57(2):409-24. DOI: 10.1016/0042-6822(74)90181-0. View

15.

Valentine R, Pereira H . Antigens and structure of the adenovirus. J Mol Biol. 1965; 13(1):13-20. DOI: 10.1016/s0022-2836(65)80076-6. View

16.

Prage L, Pettersson U . Structural proteins of adenoviruses. VII. Purification and properties of an arginine-rich core protein from adenovirus type 2 and type 3. Virology. 1971; 45(2):364-73. DOI: 10.1016/0042-6822(71)90337-0. View

17.

Smith M, Salas M, Stanley Jr W, Wahba A, Ochoa S . Direction of reading of the genetic message. II. Proc Natl Acad Sci U S A. 1966; 55(1):141-7. PMC: 285767. DOI: 10.1073/pnas.55.1.141. View

18.

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

19.

WINTERS W, Russell W . Studies on the assembly of adenovirus in vitro. J Gen Virol. 1971; 10(2):181-94. DOI: 10.1099/0022-1317-10-2-181. View

20.

Maizel Jr J, WHITE D, Scharff M . The polypeptides of adenovirus. I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. Virology. 1968; 36(1):115-25. DOI: 10.1016/0042-6822(68)90121-9. View