In vitro cleavage of RNAI
(A) and 9S RNA (B) by the E. coli
and Synechocystis sp. Rne proteins. Internally labeled
RNAI or 9S RNA was incubated in RNase E reaction buffer for 3 and 30
min with either E. coli RNase E (EcoRne)
or Synechocystis sp. Rne (SynRne), or without protein
for 30 min (C), as a negative control. The positions of the
original substrates (RNAI and 9S RNA) and their cleavage products (RNA
I-5 and 5S rRNA (5S), respectively) are indicated.
(C) Protein blots probed with RNAI and 9S RNA. Samples of
the Synechocystis sp. Rne protein (Syn) and the
E. coli RNA degradosome (Eco) were resolved by
SDS/PAGE, electroblotted onto poly(vinylidene difluoride) membrane,
and probed with 32P-labeled RNAI or 9S RNA as described in
Materials and Methods. The Synechocystis
sp. Rne (SynRne) and E. coli Rne (EcoRne)
proteins complexed with RNA are indicated.