A protein shuttle system to target RNA into mitochondria

Nucleic Acids Research, 2011, Vol. 39, No. 14, Pages e96, https://doi.org/10.1093/nar/gkr380

In Figure 2B, ‘T’ designates radiolabeled mature tRNA transcripts used as size marker. The authors do not recall whether tRNA^Ala or tRNA^His was used. However, as these two mature tRNA transcripts are 75–76 nt in size and migrate the same way in denaturing gel, it does not affect the result and conclusion of the figure. In the figure legend ‘32P-labeled in vitro-transcribed tRNA^Ala as a size marker’ should be ‘32P-labeled in vitro-transcribed mature tRNA as a size marker’.

Figures 1A, 1B and S4 contain undisclosed splicing. The raw data and a new Figure 1 are provided below. A new Figure S4 is available in the Supplementary Data. The original data for Figure 1A could not be recovered, the results of a repeat experiment are provided instead.

Figure 1A.

raw data.

Figure 1B.

raw data.

New Figure 1.

pDHFR increases tRNA import into isolated potato mitochondria. (A) On the left: ³²P-labeled in vitro-transcribed tRNA^Ala was incubated with isolated potato mitochondria (25) in the absence (—) or presence (+) of 35 pmol of pDHFR. Following standard import conditions, RNase treatment was performed either on mitochondria (Mi) or on mitoplasts (Mtp). RNAs were fractionated on a denaturing polyacrylamide gel. Equivalent loading was checked by ethidium bromide staining prior to autoradiography visualization (4 h exposure). On the right: visualization under confocal microscope of Alexa Fluor-labeled in vitro transcribed tRNA^Ala incubated with isolated potato mitochondria in the absence (2) or presence (3 and 4) of 35 pmol of pDHFR. Visualization was performed after 5 min (3) or 25 min (2 and 4) of incubation. Alexa Fluor-labeled in vitro transcribed tRNA^Ala in import medium without mitochondria was used as a control (1). (B) 32P-labeled in vitro-transcribed tRNA^Ala was incubated with isolated potato mitochondria in the absence (—) or presence (70 and 140 pmol) of pDHFR or DHFR. Methotrexate (MTX, 50 nM), or antibodies against VDAC or TOM20 (25) were added to the standard import mixture and incubated for 10 min before adding the labelled tRNA^Ala. in: 10% of input RNA (2 fmol).

SUPPLEMENTARY DATA

Supplementary Data are available at NAR Online.