Fig. 1.
Fig. 1. Characterization of the rat and mouse platelet GPV genes. (A) Cloning strategy and structure of the rat and mouse GPV genes and comparison to the human gene. The rat GPV gene was obtained by screening of a λ Dash genomic library (λVR1 clone) with a human genomic DNA probe, PCR amplification of rat genomic DNA, and rescreening of the genomic library (λVR2 clone) with the PCR fragment. The reported sequence corresponds to the λVR2 clone and contains the entire coding sequence and promoter region. The mouse GPV gene (λVM1 clone) was obtained by screening a mouse genomic library in the λ Dash vector with the rat λVR1 clone. Each clone was sequenced at least once on both strands. The human GPV gene structure16 is given for comparison. The three genes have a single intron dividing the genes in two exons. The clones and intron sizes are indicated in nucleotides (nt). Stippled, closed, and open boxes represent the coding, 5′ and 3′ untranslated, and promoter regions, respectively. The ATG start and TGA stop codons are marked with (▴). The (▵) indicates the position of a putative transcription start site. Sites for EcoRI, Xho I, HindIII, Pst I, and BamHI restriction enzymes are indicated. The EcoRI site in parentheses is a cloning site not present in the gene. (B) Comparison of human (H), rat (R), and mouse (M) GPV promoter sequences. Approximatly 500 nt of the 5′ flanking sequences upstream of the intron are shown. Nucleotide numbering on the left corresponds to Lanza et al16 for human GPV and to the clones λRV2 and λMV1 for rat and mouse GPV, respectively. A putative initiation start site is boxed. Negative nucleotide numbering on the basis of the proposed initiation site is indicated on the right. Identity with the human sequence is indicated by a (:), while hyphens denote gaps inserted to optimize the alignment. Putative binding sites for TATA, GATA, Ets, PuF, CACC, NFE-2, and the platelet specific PMS-E transcription factors are indicated by bold-underlined characters. (C) Transcriptional start sites in megakaryocyte and platelet expressed genes. The sequences flanking the +1 position for mouse terminal deoxynucleotidyl transferase (TdT) and human αIIb and α2 integrins, GPIX, and PF4 genes are compared with 5′ flanking sequences of H, R, and M GPV genes. Sites of transcription initiation for promoters lacking TATA and CAAT boxes have a consensus sequence with CA at −1, +1 (boxed) and pyrimidine-rich segments on either side (underlined).

Characterization of the rat and mouse platelet GPV genes. (A) Cloning strategy and structure of the rat and mouse GPV genes and comparison to the human gene. The rat GPV gene was obtained by screening of a λ Dash genomic library (λVR1 clone) with a human genomic DNA probe, PCR amplification of rat genomic DNA, and rescreening of the genomic library (λVR2 clone) with the PCR fragment. The reported sequence corresponds to the λVR2 clone and contains the entire coding sequence and promoter region. The mouse GPV gene (λVM1 clone) was obtained by screening a mouse genomic library in the λ Dash vector with the rat λVR1 clone. Each clone was sequenced at least once on both strands. The human GPV gene structure16 is given for comparison. The three genes have a single intron dividing the genes in two exons. The clones and intron sizes are indicated in nucleotides (nt). Stippled, closed, and open boxes represent the coding, 5′ and 3′ untranslated, and promoter regions, respectively. The ATG start and TGA stop codons are marked with (▴). The (▵) indicates the position of a putative transcription start site. Sites for EcoRI, Xho I, HindIII, Pst I, and BamHI restriction enzymes are indicated. The EcoRI site in parentheses is a cloning site not present in the gene. (B) Comparison of human (H), rat (R), and mouse (M) GPV promoter sequences. Approximatly 500 nt of the 5′ flanking sequences upstream of the intron are shown. Nucleotide numbering on the left corresponds to Lanza et al16 for human GPV and to the clones λRV2 and λMV1 for rat and mouse GPV, respectively. A putative initiation start site is boxed. Negative nucleotide numbering on the basis of the proposed initiation site is indicated on the right. Identity with the human sequence is indicated by a (:), while hyphens denote gaps inserted to optimize the alignment. Putative binding sites for TATA, GATA, Ets, PuF, CACC, NFE-2, and the platelet specific PMS-E transcription factors are indicated by bold-underlined characters. (C) Transcriptional start sites in megakaryocyte and platelet expressed genes. The sequences flanking the +1 position for mouse terminal deoxynucleotidyl transferase (TdT) and human αIIb and α2 integrins, GPIX, and PF4 genes are compared with 5′ flanking sequences of H, R, and M GPV genes. Sites of transcription initiation for promoters lacking TATA and CAAT boxes have a consensus sequence with CA at −1, +1 (boxed) and pyrimidine-rich segments on either side (underlined).

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