TY - JOUR
T1 - Identification of mitochondrial gene products by DNA-directed protein synthesis in vitro
AU - Moorman, A. F.M.
AU - Grivell, L. A.
AU - Lamie, F.
AU - Smits, H. L.
N1 - Funding Information: We are grateful to Professor P. Borst and Dr. G.S.P. Groot for helpful discussion, to Mr. G.J.B. van Ommen for help with the experiments presented in Fig. 2, to Mr. W. van Raamsdonk and Mr. C.W. Pool for much immunological advice to Mrs. N. van Harten-Loosbroek, Mrs. C. Heyting and Mr. C.F. van Kreijl for gifts of yeast submitochondrial particles, petite DNA and RNA polymerase, respectively, to Mrs. G. Reymer-van Dam for technical assistance and to those mentioned in Methods who generously provided us with antigens and antisera. This work was supported in part by a grant to P. Borst from The Netherlands Foundation for Chemical Research (S.O.N.) with financial aid from The Netherlands Organization for the Advancement of Pure Research (Z.W.O.).
PY - 1978/4/27
Y1 - 1978/4/27
N2 - 1. 1. A cell-free system, derived from Escherichia coli is highly active in the linked transcription-translation of yeast mtDNA from both wild-type and petite strains. 2. 2. The products of synthesis are short (Mr < 10 000) hydrophobic polypeptides, which show a high tendency to aggregate in a specific fashion with E. coli and mitochondrial proteins. Aggregation is extremely persistent: alkali, sodium dodecyl sulphate urea, guanidinium · HCl and carboxymethylation reduce it, but do not eliminate it completely. 3. 3. Nevertheless, results of indirect immunoprecipitation tests suggest that antigenic determinants of cytochrome c oxidase are among the products synthesized. The immunoprecipitation appears specific by criteria including competition experiments and its absence when mtDNA from low complexity petites, retaining only the gene for 21 S rRNA and some flanking sequences, is used to programme protein synthesis. Electrophoretic analysis of material precipitated by anti-cytochrome c oxidase sera reveals four discrete polypeptides with molecular weights of 7400, 6400, 5000 and 4100, which probably represent polypeptide fragments carrying antigenic determinants of cytochrome c oxidase .
AB - 1. 1. A cell-free system, derived from Escherichia coli is highly active in the linked transcription-translation of yeast mtDNA from both wild-type and petite strains. 2. 2. The products of synthesis are short (Mr < 10 000) hydrophobic polypeptides, which show a high tendency to aggregate in a specific fashion with E. coli and mitochondrial proteins. Aggregation is extremely persistent: alkali, sodium dodecyl sulphate urea, guanidinium · HCl and carboxymethylation reduce it, but do not eliminate it completely. 3. 3. Nevertheless, results of indirect immunoprecipitation tests suggest that antigenic determinants of cytochrome c oxidase are among the products synthesized. The immunoprecipitation appears specific by criteria including competition experiments and its absence when mtDNA from low complexity petites, retaining only the gene for 21 S rRNA and some flanking sequences, is used to programme protein synthesis. Electrophoretic analysis of material precipitated by anti-cytochrome c oxidase sera reveals four discrete polypeptides with molecular weights of 7400, 6400, 5000 and 4100, which probably represent polypeptide fragments carrying antigenic determinants of cytochrome c oxidase .
UR - http://www.scopus.com/inward/record.url?scp=0017813589&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/0005-2787(78)90192-2
DO - https://doi.org/10.1016/0005-2787(78)90192-2
M3 - Article
C2 - 207328
SN - 0005-2787
VL - 518
SP - 351
EP - 365
JO - BBA Section Nucleic Acids And Protein Synthesis
JF - BBA Section Nucleic Acids And Protein Synthesis
IS - 2
ER -