Cloning of cDNAs encoding human caldesmons.
Caldesmon (CDM) is a potential actomyosin regulatory protein found in smooth muscle and nonmuscle cells. Domain mapping and physical studies suggest that CDM is an elongated molecule with an N-terminal myosin/calmodulin-binding domain and a C-terminal tropomyosin/actin/calmodulin-binding domain separated by a 40-nm-long central helix. An 1100-nucleotide (nt) cDNA probe encoding the C terminus of avian caldesmon (aCDM) was used to screen a human aorta library and clone smooth-muscle and non-muscle CDM-encoding cDNAs (CDM). The human (h) smooth-muscle hCDM is 3050-3630 nt long, having variation in length in the 3'-untranslated region. The predicted hCDM protein has a high degree of identity, greater than 90%, to aCDM in the N- and C-terminal-binding domains. The central helical domain is more variable, but retains characteristic repeated peptides and an 'i, i + 4' acidic/basic amino acid (aa) motif found in aCDM which can form intra-helical salt bridges to stabilize the central helix. The predicted smooth-muscle protein is 793 aa long (93,262 Da) with a calculated pI of 5.75. As is the case for the chicken, nonmuscle hCDM is missing the central helical domain, 256 aa overall. Our nonmuscle clone is not full length, but the C-terminal end is identical to the smooth-muscle form. If the N-terminal domain is identical, as it is in the chicken, the predicted protein is 537 aa (62,558 Da). Examination of the 'junctions' at either end of the deleted central domain gives a clear indication of the splice sites and suggests that the nonmuscle form is generated by exon skipping.(ABSTRACT TRUNCATED AT 250 WORDS)
Humphrey
M B
MB
Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030.
Herrera-Sosa
H
H
Gonzalez
G
G
Lee
R
R
Bryan
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002148:Calmodulin-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000737:chemistry; D011487:Protein Conformation
D002148:Calmodulin-Binding Proteins; D004247:DNA