Michel, Rainer; Neupert, Walter
Mitochondrial Translation Products before and after Integration into the Mitochondrial Membrane in Neurospora crassa.
In: European Journal of Biochemistry, Vol. 36, Nr. 1: S. 53-67
Nascent translation products on mitochondrial ribosomes were selectively labeled in vivo in the presence of cycloheximide with radioactive leucine. They were isolated together with the ribosomes.
The labeled polypeptides show a high tendency to aggregate and can only be kept in solution in the presence of detergents such as dodecylsulfate. Also, mitochondrial ribosomes carrying nascent peptide chains easily form aggregates.
The polypeptides adhering to mitochondrial monomeric ribosomes differ from those adhering to polymeric ribosomes. Gel electrophoresis in the presence if dodecylsulfate shows for the peptidyl transfer RNA products at the monomer, an apparent molecular weight of 27000. After removing the transfer RNA, an apparent molecular weight of less than 10000 is registered. The peptides adhering to mitochondrial polymeric ribosomes display a broad range of apparent molecular weights. In contrast, translation products associated with cytoplasmic monomeric and polymeric ribosomes all show quite disperse molecular weights.
Using gel-chromatographic analysis no difference in the elution characteristics between translation products associated with mitochondrial monomeric and polymeric ribosomes was found. In both cases apparent molecular weights of about 11000 were obtained.
A kinetic study of the appearance of mitochondrial translation products in the mitochondrial membrane was carried out. A conversion process of products with lower apparent molecular weights to those with higher apparent molecular weights is observed. This suggests that mitochondrial ribosomes form polypeptides which are modified during or after integration into the membrane.
The hypothesis is discussed that mitochondria possess their own system of transcription and translation, because the hydrophobic nature of the translation products makes it necessary that they are formed inside the inner mitochondrial membrane, into which they are integrated.