Analysis of sense and naturally occurring antisense transcripts of myosin heavy chain in the human myocardium.
Naturally occurring antisense RNA has the potential to form a duplex with its complementary sense mRNA, thereby regulating protein expression. Previously, we demonstrated considerable amounts of endogenous antisense RNA for both alpha- and beta-myosin heavy chain (MHC) in rat heart suggesting a role in posttranscriptional MHC-regulation (Luther et al. [1997] J Mol Cell Cardiol 29(1):27-35). To evaluate whether antisense RNA is also involved in MHC regulation in human heart we analyzed ventricular myocardium transcripts in nonfailing hearts (n=3) and hearts from patients undergoing heart transplantation (n=5). Investigation of RNA by reverse transcription polymerase chain reaction (RT-PCR) detected an antisense RNA transcript for beta-MHC but none for alpha-MHC. Northern blot analysis of normal and failing hearts detected sense mRNA for beta-MHC, but not alpha-MHC suggesting no functionally relevant levels of alpha-MHC mRNA exist in the human ventricle. The results describe-for the first time-the existence of endogenous polyadenylated MHC antisense transcripts in the human heart. The potential effect of attenuating translation was shown in an in vitro translation assay using a synthetic antisense-oligonucleotide derived from the sequence of the naturally occurring antisense RNA.
Luther
H P
HP
Medical Clinic I, Department of Cardiology, Humboldt-University (Charité), Ziegstr. 5-9, D10117 Berlin, Germany.
Podlowski
S
S
Hetzer
R
R
Baumann
G
G
D000293:Adolescent; D000328:Adult; D015152:Blotting, Northern; D002648:Child; D004305:Dose-Response Relationship, Drug; D005260:Female; D016027:Heart Transplantation; D006801:Humans; D008163:Luminescent Measurements; D008297:Male; D008875:Middle Aged; D009206:Myocardium / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011061:Poly A / Q000378:metabolism; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D011061:Poly A; D018995:Myosin Heavy Chains