73
TABLE 10
RNA POLYMERASE ACTIVITY SOLUBILIZED FROM
MITOCHONDRIA
Treatment of
Mitochondria
in Lysis Buffer
RNA Polymerase
(units/g wet
30KS
Activity
wt.)
30KP
+ Water
4.2
69
+ 0.5 M KC1
4.2
63
+0.8% NP-40
45.5
46
+0.8% Triton X-100
34.0
37
+ 0.8% NP-40 + 0.5 M KC1
56.0
0
+ 0.8% Triton + 0.5 M KC1
68.0
15
+ Water (Sonication)
0
28
Mitochondria purified by DNase treatment were suspended in lysis buffer
(lg wet wt/10 ml). Mitochondrial samples of 1.0 ml each were then added
to an equal volume of the indicated lysis mixture which contained 0.01 M
EDTA plus additions shown. The mixtures were mixed gently for 15 min. at
4C and then centrifuged for 90 min. at 30,000 rpm (Ti 50 rotor). The
supernatants and pellets were dialyzed against TGMED Buffer for 4 hrs.
before analyzing them for RNA polymerase activity as described in the
methods. The concentration of ^H-UTP was 0.016 mM with a specific radio
activity of 2 Ci/mmole.