Novel alternatively spliced variant form of human CDK5RAP2

Figures & data

Figure 1 The human full length CDK5RAP2 (hCDK5RAP2) and its spliced variant form (hCDK5RAP2-V1). (A) Schematic diagram of human chromosome 9q33.2 minus strand from nucleotide 123151147 to nucleotide 123342437 [adapted from UCSC Genome Browser on Human Feb 2009 (GRCh37/hg19) Assembly]. Vertical bars with numbers indicate exons for hCDK5RAP2 and hCDK5RAP2-V1. Note that CDK5RAP2 exon 32 is missing in hCDK5RAP2-V1 (asterisk) due to alternative splicing. (B) Sequence alignment of hCDK5RAP2 with hCDK5RAP2-V1. Sequences were aligned by CLUSTAL 2.0.12 multiple sequence alignment software; amino acid numbers are indicated on either side. The identical sequences of hCDK5RAP2 and hCDK5RAP2-V1 are shown. The broken line corresponds to the missing amino acid sequence in hCDK5RAP2-V1. The codon (GAT) encoding aspartic acid (D, shaded in grey) for hCDK5RAP2 and hCDK5RAP2-V1 is generated from the combination of nucleotides (G-AT) between exon 32 and exon 33, and between exon 31 and exon 32, respectively. (C) Primary structure of hCDK5RAP2 and hCDK5RAP2-V1. hCDK5RAP2 encodes 1,893 amino acids (215 kDa) while hCDK5RAP2-V1 encodes 1,814 amino acids (206 kDa) lacking 79 amino acids from amino acid_1,576 to amino acid_1,654. hCDK5RAP2 (genbank accession no. NP_060719) and hCDK5RAP2-V1 (genbank accession no. NP_001011649) are shown. (D) Tissue distribution of hCDK5RAP2 and hCDK5RAP2-V1 transcripts by PCR using human cDNAs as template and the following primers: forward primer (TGG AAC GGC AAG GAT CTG AA) and reverse primer (TCA CTG CCT GGG AGG AAT CA). hCDK5RAP2 and hCDK5RAP2-V1 transcripts produce 762 bp and 525 bp PCR fragments. RPL13A was used as an internal control: forward primer (CCTG GAG GAG AAG AGG AAA GAG A) and reverse primer (TTG AGG ACC TCT GTG TAT TTG TCA A). No template was used for the negative control. Levels of hCDK5RAP2-V1 transcript were determined by densitometric analysis using the Image J software. Asterisk indicates a non-specific PCR product that was confirmed by sequence analysis.