Only the protein encoded by BC1G_01003 (called Bhl1, for ‘ B otry

Only the protein encoded by BC1G_01003 (called Bhl1, for ‘ B otrytis h ydrophobin- l ike’), showed a hydrophobicity similar to Bhp1. However, the cysteine spacing of Bhl1 differs somewhat from that of confirmed class I hydrophobins [16] (Table 1), it has a distinct hydropathy profile (additional file 2 : Figure S1), and it lacks homology to other fungal hydrophobins (data not shown). Table 1 Sequence characteristics of B. cinerea

hydrophobins and hydrophobin-like proteins. Name/predicted class Size Spacing of cysteine residues GRAVY Bhp1 (BC1G_15273) 111/93 N- 34-C- AP24534 datasheet 7 -CC- 18 -C- 15 -C- 5 -CC- 17 -C- 7 0.57 Consensus spacing class I   N- Xn-C- (5-8) -CC-(17-39) -C-(8-23) -C-(5-6) -CC-(6-18) -C-(2-13)   Bhp2 (BC1G_03994) 98/77 N- 33-C- 6 -CC- 11 -C- 16 -C- 8 -CC- 10 -C- 6 0.42 Bhp3 (BC1G_01012) 98/80 N- 34-C- 8 -CC- 11 -C- 16 -C- 8 -CC- 10 -C- 3 0.30 Consensus spacing class II   N- Xn-C-(9-10) -CC- 11 -C- 16 -C-(6-9) -CC- 10 -C- (3-7)   Bhl1 (BC1G_01003) 145/125 N- 60-C- 9 -CC- 31 -C- 8 -C-

7 -CC- 16 -C- 6 0.76 BC1G_02483 234/211 N- 82-C- 8 -CC- 7 -C- 5 -C- 9 -CC- 8 -C- 107 -0.10 BC1G_03277 178/160 N-111-C- 7 -CC- 10 -C- 17 -C- 8 -CC- 12 -C- 5 -0.43 BC1G_04521 181/157 N-120-C- 7 -CC- 10 -C- 10 -C- 9 -CC- 4 -C- 13 0.01 BC1G_11117 109/88 N- 35-C- 10 -CC- 15 -C- 18 -C- 8 -CC- 11 -C- 4 -0.77 BC1G_12747 106/86 N- 37-C- 3 -CC- 10 -C- 13 -C- 18 -CC- 4 -C- 13 -0.28 For the three hydrophobins Bhp1 (class I), Bhp2 and Bhp3 (both class II), and for six hydrophobin-like proteins, the cysteine spacing is shown. PD0332991 in vitro Consensus

cysteine spacings for class I and class II proteins were taken from [16]. The sizes (amino acids) of the unprocessed and processed Tryptophan synthase proteins are indicated. N: N-terminus; Xn: Undefined number of amino acids; Underlined: Strictly conserved spacing; GRAVY: Grand average of hydropathicity of the region covering the eight cysteines. Positive GRAVY values indicate hydrophobicity [53]. Bhp1 is 111 amino acids long and contains eight cysteines with spacing as described for the class I hydrophobin consensus sequence [16]. It shows 30% identity to Xph1 of the lichen fungus Xanthoria parietina, and 29% identity to Mpg1 of Magnaporthe oryzae (Figure 1A). The hydropathy plot of Bhp1 shows similarity to that of Mpg1 and of other class I hydrophobins (Figure 1C; data not shown). Bhp2 and Bhp3 are both 98 amino acids long and 27% identical to each other. Both proteins match the consensus cysteine spacing of class II hydrophobins (Table 1) [16]. Bhp2 shares 37%, and Bhp3 29% identity with M. oryzae Mhp1 (Figure 1B). The hydropathy plots of Bhp2 and Mhp1 are similar (Figure 1D). Figure 1 Sequence alignments and hydropathy plots of B. cinerea hydrophobins and confirmed class I and II hydrophobins. A: Amino acid alignment of Bhp1 and class I hydrophobins. B: Amino acid alignment of Bhp2/3 and class II hydrophobins. The signal peptides are underlined. Hcf3 (Acc.

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