Computational and Experimental Analyses Reveal Previously Undetected Coding Exons of the KRIT1 (CCM1) Gene

• Published in: Genomics (San Diego, Calif.) Calif.), 2001-01, Vol.71 (1), p.123-126
• Main Authors: Sahoo, Trilochan, Goenaga-Diaz, Eduardo, Serebriiskii, Ilya G., Thomas, James W., Kotova, Elena, Cuellar, Jacob G., Peloquin, Joanna M., Golemis, Erica, Beitinjaneh, Firas, Green, Eric D., Johnson, Eric W., Marchuk, Douglas A.
• Format: Article
• Language: English
• Subjects: Alternative Splicing; Amino Acid Sequence; Animals; Biological and medical sciences; CCM1 gene; cerebral cavernous malformation type 1 gene; DNA, Complementary - metabolism; Exons; Expressed Sequence Tags; Frameshift Mutation; Fundamental and applied biological sciences. Psychology; Genes. Genome; Humans; KRIT1 gene; KRIT1 Protein; Mice; Microtubule-Associated Proteins; Models, Genetic; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Mutation; Proto-Oncogene Proteins - chemistry; Proto-Oncogene Proteins - genetics; Reverse Transcriptase Polymerase Chain Reaction; Sequence Homology, Amino Acid; Software; start codons
• ISSN: 0888-7543; 1089-8646
• DOI: 10.1006/geno.2000.6426

A notable difficulty in annotating genomic sequence is identifying the correct 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 putative exons upstream of the previously described first exon. These additional 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 presence 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 establish 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 and comparative sequence analysis for refining gene structure.