Measuring mRNA decay in human mitochondria.
Human mitochondria contain a genome encoding 13 proteins, all of which are components of respiratory chain complexes. Mutations in human mitochondrial DNA often have pathological consequences. Although 12 of the mitochondrial mRNAs are generated from the same polycistronic transcript, the steady-state level of each mRNA differs. The stability of each mitochondrial mRNA is post-transcriptionally controlled by polyadenylation and deadenylation. However, the molecular mechanism by which each mRNA attains a unique stability is not fully understood. In this report, we describe a practical method for measuring the half-lives of human mitochondrial mRNAs using quantitative real-time reverse transcription PCR.
Nagao
Asuteka
A
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Tokyo, Japan.
Hino-Shigi
Narumi
N
Suzuki
Tsutomu
T
D001483:Base Sequence; D017931:DNA Primers; D004996:Ethidium / Q000378:metabolism; D006207:Half-Life; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000235:genetics*; D011061:Poly A / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D012333:RNA, Messenger; D011061:Poly A; D004996:Ethidium