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Vol. 15, Issue 3, 1211-1223, March 2004

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Role of Binding of Plectin to the Integrin 4 Subunit in the Assembly of Hemidesmosomes

The Netherlands Cancer Institute, Division of Cell Biology, 1066 CX Amsterdam, The Netherlands

Submitted September 25, 2003; Revised November 20, 2003; Accepted November 24, 2003
Monitoring Editor: Richard Hynes

We have previously shown that plectin is recruited into hemidesmosomes through association of its actin-binding domain (ABD) with the first pair of fibronectin type III (FNIII) repeats and a small part of the connecting segment (residues 1328–1355) of the integrin 4 subunit. Here, we show that two proline residues (P1330 and P1333) in this region of the connecting segment are critical for supporting 4-mediated recruitment of plectin. Additional binding sites for the plakin domain of plectin on 4 were identified in biochemical and yeast two-hybrid assays. These sites are located at the end of the connecting segment (residues 1383–1436) and in the region containing the fourth FNIII repeat and the C-tail (residues 1570–1752). However, in cells, these additional binding sites cannot induce the assembly of hemidesmosomes without the interaction of the plectin-ABD with 4. Because the additional plectin binding sites overlap with sequences that mediate an intramolecular association of the 4 cytoplasmic domain, we propose that they are not accessible for binding and need to become exposed as the result of the binding of the plectin-ABD to 4. Furthermore, these additional binding sites might be necessary to position the 4 cytoplasmic domain for an optimal interaction with other hemidesmosomal components, thereby increasing the efficiency of hemidesmosome assembly.

Abbreviations used: ABD, actin-binding domain; BP, bullous pemphigoid; CS, connecting segment; FNIII, fibronectin type III repeat; HD, hemidesmosome; IL2R, interleukin 2 receptor; MD-EBS, muscular dystrophy associated with epidermolysis bullosa simplex; PA-JEB, pyloric atresia associated with junctional epidermolysis bullosa; PBS, plectin binding site.

^* Corresponding author. E-mail address: a.sonnenberg{at}nki.nl.

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