N u c l e a r s k e l e t o n structures in s o m e n o r m a l and t u m o r cells

I. B. Zbarsky
Laboratory of Biochemistry, N.K. Koltzov Institute of Developmental Biology, Academy of Sciences of the
USSR, Vavilow Street 26, 117808 GSP-1, Moscow 117334, USSR

Abstract

Nuclear protein fractions, described earlier, were identified as constituents of the nuclear sap (the'globulin
fraction'), that of the nucleoli and ribonucleoprotein network (the 'acidic protein'), and of the nuclear
envelope (the 'residual protein'). The latter two fractions compose the protein skeleton of the cell nucleus.
An essential difference between electrophoretic profiles of nuclear skeleton structures in experimental
tumors and those of normal tissues was revealed. T u m o r preparations contained more high molecular weight
polypeptides and, in earlier stages of growth, low molecular weight components as well. Fractionation of the
nuclear matrix proteins showed that the bulk of them are soluble in diluted alkali. The alkali-insoluble
fraction retains the shape of the nucleus and appears in the electron microscope as a spongy nuclear skeleton.
A finely dispersed fraction sedimenting from the alkaline suspension is enriched with the pore complexes. The
fractions obtained differ in protein composition and probably contain protein components which are similar
in molecular weights but non-identical.

Introduction laboratory (16).

In our earlier studies a deoxyribonucleoprotein
fraction soluble in 2 M NaC1, an 'acidic protein'
fraction soluble in 0.02 N N a O H , and a fraction of a
'residual protein' insoluble in diluted alkali were
obtained by fractionation of isolated cell nuclei
(41). In various t u m o r tissues the content of the
residual protein fraction and usually likewise of the
acidic protein were found to be considerably
enlarged (12, 38, 39), the proportion of the residual
protein being increased in the course of the growth
of the transplantable t u m o r (31). Similar fractions
were isolated by other workers (11, 21, 33, 36). The
origin of RP from the nuclear envelope has been
reported previously (33) and was confirmed in our
Abbreviations: KD - kilodaltons; NM nuclear matrix; PAAG
polyacrylamide gel; PC - pore complex; RP residual protein.
To our scheme of fractionation we added a previ-
ous extraction with physiological saline and studied
frozen thin sections and isolated nuclei in the light
microscope after each extraction. These studies
have shown that the 'globulin fraction' cytologically
corresponded to the nuclear sap, deoxyribonucleo-
protein fraction - to chromatin, acidic protein - to
nucleoli and ribonucleoprotein network, and the
RP - to the nuclear envelope (16, 44). These results
were confirmed by electron microscopy. It was
shown that apart from the normal c o m p o n e n t of
the RP corresponding to the nuclear envelope, in
Walker carcinosarcoma the preparation included
an amorphous component characteristic of tumors
and partly extractable with aqueous butanol (40,
43, 46). This finding has substantiated our earlier
data on the presence in t u m o r RP of a particular
tryptophan-rich fraction extractable with organic
solvents (39, 42).

Molec. Biol. Rep. 7, 139-148 (1981). 0301 4851/81/0073-0139 $2.00.

9 Dr W. Junk Publishers, The Hague. Printed in The Netherlands.