Isolation and Characterization of Human Fast Skeletal β Troponin T cDNA: Comparative Sequence Analysis of Isoforms and Insight into the Evolution of Members of a Multigene Family

A cDNA encoding human fast skeletal β troponin T (β TnT_f) has been isolated and characterized from a fetal skeletal muscle library. The cDNA insert is 1,000 bp in length and contains the entire coding region of 777 bp and 5′ and 3′ untranslated (UT) segments of 12 and 211 bp, respectively. The 3′ UT segment shows the predicted stem-loop structure typical of eukaryotic mRNAs. The cDNA-derived amino acid sequence is the first available sequence for human β TnT_f protein. It is encoded by a single-copy gene that is expressed in a tissue-specific manner in fetal and adult fast skeletal muscles. Although the human β TnT_f represents the major fetal isoform, the sequence information indicates that this cDNA and the coded protein are quite distinct from the fetal and neonatal TnT_f isoforms reported in other mammalian fetal muscles. The hydropathy plot indicates that human β TnT_f is highly hydrophilic along its entire length. The protein has an extremely high degree of predicted α-helical content involving the entire molecule except the carboxy-terminal 30 residues. Comparative sequence analysis reveals that the human β TnT_f shares a high level of sequence similarity in the coding region with other vertebrate TnT_f and considerably reduced similarity with slow skeletal and cardiac TnT cDNAs. The TnT isoforms have a large central region consisting of amino acid residues 46–204 which shows a high sequence conservation both at the nucleotide and amino acid levels. This conserved region is flanked by the variable carboxy-terminal and an extremely variable amino-terminal segment. The tropomyosin-binding peptide of TnT, which is represented by amino acid residues 47–151 and also includes a part of troponin I binding region, is an important domain of this central segment. It is suggested that this conserved segment is encoded by an ancestral gene. The variable regions of vertebrate striated TnT isoforms reflect the subsequent addition and modification of genomic sequences to give rise to members of the TnT multigene family.