Supplemental data for “A Cell Plate–Specific Callose Synthase and Its Interaction with Phragmoplastin”

 

Z. Hong, A.J. Delauney, and D. P.S. Verma

 

The following data supplements the information provided in Figure 3 (Hong, Delauney, and Verma; 2001). Protein sequences of the Arabidopsis callose synthase catalytic subunit were deduced from cDNA (CalS1) and genomic DNA sequences (CalS2-12).  The coding region of each sequence was compiled based on the CalS1 sequence, which was determined experimentally.  CalS11-12 sequences are from GenBank while CalS9 is from the NCBI protein database. Other CalS sequences were deduced as follows: Exons of each sequence were first identified using the Procrustes software program (http://hto13.usc.edu/software/procrustes) with CalS1 as a target peptide sequence.  CalS1 protein sequence was compared, using TblastN (http://www.ncbi.nlm.nih.gov/blast), with genomic DNA translated into three reading frames.  This resulted in identification of more exons than indicated in the corresponding GenBank data files.  Annotation of genomic DNA sequences was finalized by combining exons that best fit with the experimentally determined CalS1 sequence and other known 1,3-b-glucan synthase sequences.  Similar analysis has been done on this group of proteins refered as GSL (glucan synthase-like) by C. Somerville’s group (http://cellwall.stanford.edu/gsl/arabidopsis/structure.shtml).  The phylogenetic relationship (Figure 3B) was established using the deduced CalS protein sequences.  The CalS family contains two types of genes, CalS1-10 fall into one group of large genes containing up to 50 exons while CalS11-12 contain 2-3 exons.  All together, CalS sequences comprise the longest coding DNAs for any known plant protein.

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