PMID- 10446244
OWN - NLM
STAT- MEDLINE
DCOM- 19991021
LR  - 20211203
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 27
IP  - 17
DP  - 1999 Sep 1
TI  - Visualization of double-stranded RNAs from the myotonic dystrophy protein kinase 
      gene and interactions with CUG-binding protein.
PG  - 3534-42
AB  - Myotonic dystrophy (DM) is associated with a (CTG) (n) triplet repeat expansion 
      in the 3'-untranslated region of the myotonic dystrophy protein kinase (DMPK) 
      gene. Using electron microscopy, we visualized large RNAs containing up to 130 
      CUG repeats and studied the binding of purified CUG-binding protein (CUG-BP) to 
      these RNAs. Electron microscopic examination revealed perfect double-stranded 
      (ds)RNA segments whose lengths were that expected for duplex RNA. The RNA 
      dominant mutation model for DM pathogenesis predicts that the expansion mutation 
      acts at the RNA level by forming long dsRNAs that sequester certain RNA-binding 
      proteins. To test this model, we examined the subcellular distribution and 
      RNA-binding properties of CUG-BP. While previous studies have demonstrated that 
      mutant DMPK transcripts accumu-late in nuclear foci, the localization pattern of 
      CUG-BP in both normal and DM cells was similar. Although CUG-BP in nuclear 
      extracts preferentially photocrosslinked to DMPK transcripts, this binding was 
      not proportional to (CUG) (n) repeat size. Moreover, CUG-BP localized to the base 
      of the RNA hairpin and not along the stem, as visualized by electron micro-scopy. 
      These results provide the first visual evidence that the DM expansion forms an 
      RNA hairpin structure and suggest that CUG-BP is unlikely to be a sequestered 
      factor.
FAU - Michalowski, S
AU  - Michalowski S
AD  - Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel 
      Hill, Chapel Hill, NC 27599, USA.
FAU - Miller, J W
AU  - Miller JW
FAU - Urbinati, C R
AU  - Urbinati CR
FAU - Paliouras, M
AU  - Paliouras M
FAU - Swanson, M S
AU  - Swanson MS
FAU - Griffith, J
AU  - Griffith J
LA  - eng
GR  - GM31819/GM/NIGMS NIH HHS/United States
GR  - T32 AI07110-18/AI/NIAID NIH HHS/United States
GR  - T32 CA09156-24/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (3' Untranslated Regions)
RN  - 0 (CELF1 Protein)
RN  - 0 (CELF1 protein, human)
RN  - 0 (DMPK protein, human)
RN  - 0 (RNA, Double-Stranded)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Ribonucleoproteins)
RN  - EC 2.7.11.1 (Myotonin-Protein Kinase)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
SB  - IM
MH  - 3' Untranslated Regions/genetics
MH  - CELF1 Protein
MH  - Cells, Cultured
MH  - Fibroblasts/metabolism
MH  - Gene Library
MH  - HeLa Cells
MH  - Humans
MH  - Microscopy, Electron
MH  - Models, Genetic
MH  - Myotonin-Protein Kinase
MH  - Nucleic Acid Conformation
MH  - Plasmids
MH  - Protein Binding
MH  - Protein Serine-Threonine Kinases/*genetics/*metabolism/ultrastructure
MH  - RNA, Double-Stranded/*genetics/*metabolism/ultrastructure
MH  - RNA-Binding Proteins/*genetics/*metabolism/ultrastructure
MH  - Recombinant Fusion Proteins/metabolism
MH  - Ribonucleoproteins/*genetics/*metabolism/ultrastructure
MH  - Trinucleotide Repeats/genetics
PMC - PMC148598
EDAT- 1999/08/14 00:00
MHDA- 1999/08/14 00:01
PMCR- 1999/09/01
CRDT- 1999/08/14 00:00
PHST- 1999/08/14 00:00 [pubmed]
PHST- 1999/08/14 00:01 [medline]
PHST- 1999/08/14 00:00 [entrez]
PHST- 1999/09/01 00:00 [pmc-release]
AID - gkc526 [pii]
AID - 10.1093/nar/27.17.3534 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 1999 Sep 1;27(17):3534-42. doi: 10.1093/nar/27.17.3534.