Description of the experimental result of two-hybrid screening

Features of protein-protein interaction pairs

This section describes the protein-protein interaction pairs and the featuresof the predicted protein sequence.

(1) Yeast two-hybrid screening and subsequent determinationof interacting domains.

Domain organization for bait and prey KIAA proteins are shown. The interacting domain used as bait are indicated by violet bars. Blue bars indicateindependent clones (one bar represents one prey). Length of the bluebars indicate the sequence of the KIAA protein contained within an isolatedprey.

(2) Analysis of Motifs, Domains, and Membrane-spanningregions

The predicted protein sequences were examined for motifs present in the InterPro database. Because weakly defined sequence motifs appear too manytimes in the HUGE database and are, thus, unlikely to be informative, thefollowing motifs were excluded from the analysis: amidation site; N-glycosylationsite; cAMP- and cGMP-dependent protein kinase phosphorylation site; caseinkinase II phosphorylation site; N-myristoylation site; protein kinase Cphosphorylation site; and tyrosine kinase phosphorylation site.

Motifs/Domains in the InterPro database were searched for by InterProScan. (Zdobnov, EM, and Apweiler, R. InterProScan--an integration platform for the signature-recognition methods in InterPro" Bioinformatics 2001; 17:847-848).

Membrane-spanning region were predicted by SOSUI (Hirokawa, T., Boon-Chieng, S., Mitaku, S. "SOSUI: classification and secondary structure prediction system for membrane proteins" Bioinformatics 1998; 14:378-379).