T. Sahoo et al., Computational and experimental analyses reveal previously undetected coding exons of the KRIT1 (CCM1) gene, GENOMICS, 71(1), 2001, pp. 123-126
A notable difficulty in annotating genomic sequence is identifying the corr
ect start codon in a gene. An important such case has been found with KRIT1
, the cerebral cavernous malformation type 1 (CCM1) gene. Analysis of human
and mouse genomic sequence encompassing the region containing KRIT1/Krit1
using exon/gene-prediction and comparative alignment programs revealed puta
tive exons upstream of the previously described first exon. These additiona
l candidate exons show significant matches to mouse and human ESTs that are
contiguous with and extend upstream from the previously designated 5' end
of the KRIT1 cDNA sequence. RT-PCR and 5'RACE experiments confirm the prese
nce of four additional upstream coding exons that encode an additional 207
amino acids. Importantly, a novel frameshift mutation in one of these newly
identified KRIT1 exons has been found in a CCM1 family. These data establi
sh the authentic KRIT1 amino acid sequence and suggest that the additional
KRIT1 exons may harbor mutations in other CCM1 families. In addition, these
results provide another example of the utility of rigorous computational a
nd comparative sequence analysis for refining gene structure. (C) 2001 Acad
emic Press.