Translation Products in vitro of Mitochondrial Messenger RNA from Neurospora crassa

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Abstract
A submitochondrial polypeptide‐synthesizing system prepared from Neurospora crassa actively incorporates radioactive amino acids in vitro in the absence of added messenger RNA. Translation products were fractionated into what are most probably single polypeptide chains by filtration through Sephadex G‐100 in the presence of dodecylsulfate. The majority of incorporated amino acids is present in two partially resolved fractions of apparent molecular weights 12000 (P1) and 10000 (P2). P2 (but not P1) forms large‐molecular‐weight aggregates during concentration in the presence of 0.5% dodecylsulfate. Both P1 and P2 aggregate in the presence of 0.01% dodecylsulfate. P1 can be separated from P2 by selective extraction with 90% methanol.Purified mitochondrial DNA from Neurospora crassa was transcribed in vitro by Escherichia coli RNA polymerase. The resulting transcript was translated in a cell‐free system from E. coli into two polypeptide fractions having apparent molecular weights identical to P1 and P2.These data suggest that in Neurospora crassa the major proteins coded by mitochondrial cistrons and synthesized by endogenous mitochondrial polyribosomes are polypeptides of apparent molecular weights 12000 and 10000.

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