Overexpression of v-myb oncogene or c-myb proto-oncogene in insect cells: characterization of newly induced nucleolus-like structures accumulating Myb protein.
The oncoprotein v-Myb induces myeloid leukemia and its cellular counterpart c-Myb is involved in the regulation of hematopoiesis. Although intensively studied, their precise subcellular localization is not known. In order to expand our knowledge in this respect, we used an artificial system overexpressing these proteins. We investigated the subcellular localization of Myb proteins in cultured non-synchronized insect cells transfected with recombinant baculoviruses overexpressing either v-myb oncogene or c-myb proto-oncogene. The cell expressing Myb proteins underwent extensive nuclear changes and exhibited distinct nuclear structures resembling nucleoli. The bulk of v-Myb and c-Myb proteins accumulated in such nucleolus-like structures which, according to the nucleolar nomenclature, we classified to three types: compact of enlarged size (type I), large ring-shaped (type II) and with nucleolonemas (type III). We investigated these structures for the presence of important nucleolar macromolecules in order to establish whether they were compatible with the function in the production of ribosomes. Strikingly, our results indicated that the different forms of these structures did not represent genuine nucleoli. They rather reflected progressive changes, induced by the virus infection and high expression of v-myb genes, accompanied by the formation of these prominent nucleolus-like structures highly enriched in Myb protein. Gradual changes in number of individual nucleolus-like forms during infection, increasing amount of Myb protein and DNA localized in them together with decreasing amount of RNA and their different interaction with viral particles indicate that the nucleolus-like structure of type I is a precursor of the type II and finally of the type III.
Stokrová
Jitka
J
Department of Biopolymers Micromorphology, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 37 Praha 6, Czech Republic. stokrova@img.cas.cz
Korb
Jan
J
Pliss
Artem
A
Raska
Ivan
I
Stulík
Jan
J
Dvoráková
Martina
M
D000818:Animals; D016367:Baculoviridae; D002460:Cell Line; D002466:Cell Nucleolus / Q000378:metabolism* / Q000648:ultrastructure; D022082:Cellular Structures / Q000378:metabolism* / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis / Q000378:metabolism; D004247:DNA / Q000032:analysis / Q000235:genetics; D005455:Fluorescent Antibody Technique; D020570:Genes, myb / Q000235:genetics*; D005822:Genetic Vectors; D016253:Microscopy, Immunoelectron; D020626:Oncogene Proteins v-myb / Q000235:genetics / Q000378:metabolism*; D020598:Proto-Oncogene Proteins c-myb / Q000235:genetics / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000032:analysis / Q000235:genetics; D018411:Spodoptera
D002868:Chromosomal Proteins, Non-Histone; D020626:Oncogene Proteins v-myb; D020598:Proto-Oncogene Proteins c-myb; D012335:RNA, Ribosomal; C047458:fibrillarin; D004247:DNA