Modulation of ligand binding by alternative splicing of the alphaPS2 integrin subunit.
The Drosophila alphaPS2 integrin subunit is found in two isoforms. alphaPS2C contains 25 residues not found in alphaPS2m8, encoded by the alternative eighth exon. Previously, it was shown that cells expressing alphaPS2C spread more effectively than alphaPS2m8 cells on fragments of the ECM protein Tiggrin, and that alphaPS2C-containing integrins are relatively insensitive to depletion of Ca(2+). Using a ligand mimetic probe for Tiggrin affinity (TWOW-1), we show that the affinity of alphaPS2CbetaPS for this ligand is much higher than that of alphaPS2m8betaPS. However, the two isoforms become more similar in the presence of activating levels of Mn(2+). Modeling indicates that the exon 8-encoded residues replace the third beta strand of the third blade of the alpha subunit beta-propeller structure, and generate an exaggerated loop between this and the fourth strand. alphaPS2 subunits with the extra loop structure but with an m8-like third strand, or subunits with a C-like strand but an m8-like short loop, both fail to show alphaPS2C-like affinity for TWOW-1. Surprisingly, a single C > m8-like change at the third strand-loop transition point is sufficient to make alphaPS2C require Ca(2+) for function, despite the absence of any known cation binding site in this region. These data indicate that alternative splicing in integrin alpha subunit extracellular domains may affect ligand affinity via relatively subtle alterations in integrin conformation. These results may have relevance for vertebrate alpha6 and alpha7, which are alternatively spliced at the same site.
Bunch
Thomas A
TA
Department of Molecular and Cellular Biology, Arizona Cancer Center, 1515 N. Campbell Ave., Tucson, Arizona 85724, USA. tbunch@email.arizona.edu
Kendall
Timmy L
TL
Shakalya
Kishore
K
Mahadevan
Daruka
D
Brower
Danny L
DL
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D039001:Integrin alpha Chains / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000737:chemistry; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry; D016415:Sequence Alignment
D029721:Drosophila Proteins; D016326:Extracellular Matrix Proteins; D039001:Integrin alpha Chains; D008024:Ligands; D020033:Protein Isoforms; D021122:Protein Subunits; C089678:Tig protein, Drosophila; C082188:if protein, Drosophila; D002118:Calcium