Abstract
A rapid method for the isolation of intracytoplasmic A particles, the putative intracellular nucleoprotein cores of mouse mammary tumor virus (MTV), is presented. Spontaneous C3H/He mouse mammary tumors and transplantable mouse Leydig cell tumor were used as source material. Large aggregations of intracytoplasmic A particles were separated from cellular contaminants on discontinuous sucrose gradients and subsequently further purified by isopycnic banding in linear sucrose gradients. The purified particles were solubilized in sodium dodecyl sulfate, and the structural proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified mouse mammary tumor virions were solubilized, and their proteins were analyzed in the same way. Comparison of co-electrophoretic gels indicated a lack of correlation in the molecular size of the major structural proteins in intracytoplasmic A particles and MTV. The three major proteins of the A particles were calculated to be 80,000; 35,000; and 20,000 daltons. Five major polyacrylamide gel electrophoresis bands were obtained with purified MTV; these were 90,000; 69,000; 55,000; 37,000; and 27,500 daltons. These figures showed good correlation with those published for MTV by Nowinski et al. These results suggest the need for the reexamination of the current tenet that intracytoplasmic particles represent intracellular MTV nucleoprotein cores.
MeSH Terms
Animals
Centrifugation, Density Gradient
Cytoplasm/microbiology
Electrophoresis, Polyacrylamide Gel
Female
Inclusion Bodies, Viral
Leydig Cell Tumor/microbiology
Male
Mammary Neoplasms, Experimental/microbiology
Mammary Tumor Virus, Mouse/analysis
Mice
Mice, Inbred C3H
Microscopy, Electron
Molecular Weight
Neoplasms, Experimental/microbiology
Nucleoproteins/analysis,isolation & purification
Peptides/analysis
Sodium Dodecyl Sulfate
Sucrose
Testicular Neoplasms/microbiology
Viral Proteins/analysis,isolation & purification
Chemicals
Nucleoproteins
Peptides
Viral Proteins
Sodium Dodecyl Sulfate
Sucrose
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Smith G H
Wivel N A
References (10)
10 references, click to expand
-
Separation of subcellular components and viruses by combined rate- and isopycnic-zonal centrifugation.
Natl Cancer Inst Monogr. 1966 Jun;21:253-83
PMID: 5951397
-
Cytoplasmic inclusions in estrogen-induced testicular interstitial-cell tumors in mice.
J Natl Cancer Inst. 1967 Jul;39(1):67-74
PMID: 4165700
-
Cytochemical studies on the mouse mammary tumor virus.
Cancer Res. 1967 Nov;27(11):2179-96
PMID: 4294533
-
The extraction of intracisternal A-particles from a mouse plasma-cell tumor.
Cancer Res. 1968 Oct;28(10):2137-48
PMID: 5696942
-
[Isolation and purification of viral particles of type A present in the cytoplasm of mediastinal lymphoma induced by ethyl-urethane in Swiss mice].
C R Acad Sci Hebd Seances Acad Sci D. 1970 Aug 10;271(6):624-7
PMID: 4989921
-
[Isolation of intracytoplasmic type "A" particles from murine lymphoma induced by ethyl-urethan].
Int J Cancer. 1971 Sep 15;8(2):326-37
PMID: 4332370
-
Isolation and partial characterization of intracytoplasmic A particles.
Virology. 1972 Apr;48(1):270-4
PMID: 4335936
-
Properties of the intracytoplasmic A particles purified from mouse tumors.
Virology. 1972 Jul;49(1):61-78
PMID: 4114180
-
[Electron microscopy of virus-like inclusions in mouse mammary epitheliomas].
Bull Assoc Fr Etud Cancer. 1955;42(2):163-78
PMID: 13240241
-
PLASTIC EMBEDDING MIXTURES FOR USE IN ELECTRON MICROSCOPY.
Stain Technol. 1964 Mar;39:111-4
PMID: 14127791