PMID- 10441569
OWN - NLM
STAT- MEDLINE
DCOM- 19991004
LR  - 20181113
IS  - 0002-9297 (Print)
IS  - 0002-9297 (Linking)
VI  - 65
IP  - 3
DP  - 1999 Sep
TI  - Congenital end-plate acetylcholinesterase deficiency caused by a nonsense
      mutation and an A-->G splice-donor-site mutation at position +3 of the
      collagenlike-tail-subunit gene (COLQ): how does G at position +3 result in
      aberrant splicing?
PG  - 635-44
AB  - Congenital end-plate acetylcholinesterase (AChE) deficiency (CEAD), the cause of 
      a disabling myasthenic syndrome, arises from defects in the COLQ gene, which
      encodes the AChE triple-helical collagenlike-tail subunit that anchors catalytic 
      subunits of AChE to the synaptic basal lamina. Here we describe a patient with
      CEAD with a nonsense mutation (R315X) and a splice-donor-site mutation at
      position +3 of intron 16 (IVS16+3A-->G) of COLQ. Because both A and G are
      consensus nucleotides at the +3 position of splice-donor sites, we constructed a 
      minigene that spans exons 15-17 and harbors IVS16+3A-->G for expression in COS
      cells. We found that the mutation causes skipping of exon 16. The mutant
      splice-donor site of intron 16 harbors five discordant nucleotides (at -3, -2,
      +3, +4, and +6) that do not base-pair with U1 small-nuclear RNA (snRNA), the
      molecule responsible for splice-donor-site recognition. Versions of the minigene 
      harboring, at either +4 or +6, nucleotides complementary to U1 snRNA restore
      normal splicing. Analysis of 1,801 native splice-donor sites reveals that
      presence of a G nucleotide at +3 is associated with preferential usage, at
      positions +4 to +6, of nucleotides concordant to U1 snRNA. Analysis of 11
      disease-associated IVS+3A-->G mutations indicates that, on average, two of three 
      nucleotides at positions +4 to +6 fail to base-pair, and that the nucleotide at
      +4 never base-pairs, with U1 snRNA. We conclude that, with G at +3, normal
      splicing generally depends on the concordance that residues at +4 to +6 have with
      U1 snRNA, but other cis-acting elements may also be important in assuring the
      fidelity of splicing.
FAU - Ohno, K
AU  - Ohno K
AD  - Department of Neurology, Mayo Clinic, Rochester, MN 55905, USA.
FAU - Brengman, J M
AU  - Brengman JM
FAU - Felice, K J
AU  - Felice KJ
FAU - Cornblath, D R
AU  - Cornblath DR
FAU - Engel, A G
AU  - Engel AG
LA  - eng
GR  - NS6277/NS/NINDS NIH HHS/United States
PT  - Case Reports
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Am J Hum Genet
JT  - American journal of human genetics
JID - 0370475
RN  - 0 (Muscle Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Small Nuclear)
RN  - 9007-34-5 (Collagen)
RN  - EC 3.1.1.7 (Acetylcholinesterase)
RN  - EC 3.1.1.7 (COLQ protein, human)
SB  - IM
MH  - Acetylcholinesterase/*deficiency/*genetics/metabolism
MH  - Alternative Splicing/*genetics
MH  - Animals
MH  - Base Pairing
MH  - Base Sequence
MH  - COS Cells
MH  - *Collagen
MH  - DNA Mutational Analysis
MH  - Exons/genetics
MH  - Female
MH  - Gene Expression
MH  - Humans
MH  - Introns/genetics
MH  - Male
MH  - Middle Aged
MH  - Motor Endplate/*enzymology/physiopathology
MH  - *Muscle Proteins
MH  - *Mutation
MH  - Pedigree
MH  - RNA, Messenger/analysis/genetics
MH  - RNA, Small Nuclear/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Transfection
PMC - PMC1377969
EDAT- 1999/08/12 10:00
MHDA- 2000/03/21 09:00
CRDT- 1999/08/12 10:00
PHST- 1999/08/12 10:00 [pubmed]
PHST- 2000/03/21 09:00 [medline]
PHST- 1999/08/12 10:00 [entrez]
AID - S0002-9297(07)62314-3 [pii]
AID - 10.1086/302551 [doi]
PST - ppublish
SO  - Am J Hum Genet. 1999 Sep;65(3):635-44. doi: 10.1086/302551.