Cutin monomers, generated by the low levels of constitutively expressed cutinase, induce high levels of cutinase that can help pathogenic fungi to penetrate into the host through the cuticle whose major structural polymer is cutin. We cloned three highly homologous cutinase genes, cut 1, cut 2, and cut 3, from Fusarium solani f. pisi (Nectria hematococca). Amino acid sequence deduced from the nucleotide sequence of cut 1 and cut 2/3 matched with that of the peptides from cutinase 1 and cutinase 2, respectively, isolated from F. solani pisi grown on cutin as the sole carbon source. Induction of -glucuronidase gene fused to the promoters of the cutinases integrated into F. solani pisi genome indicates that cut 2 is constitutively expressed and induced under starvation whereas cut 1 is highly induced by cutin monomers. A palindrome binding protein (PBP) previously cloned binds only to palindrome 1 of cut 1 promoter but not palindrome 1 of cut 2/3 which contains two base substitutions. PBP is thought to interfere with the binding of CTF1, the transcription factor involved in induction, to cut 1 promoter and thus keep cut 1 gene repressed until induced by cutin monomers. Since PBP cannot bind palindrome 1 of cut 2, this gene is not repressed. CTF1 does not transactivate cut 2 promoter. A new Cys6Zn2 motif-containing transcription factor, CTF1, that binds palindrome 2 was cloned and sequenced. In yeast, CTF1 transactivates cut 2 promoter but not cut 1 promoter unless its palindrome 1 is mutated, unlike CTF1 which transactivates cut 1. Thus, CTF1 is involved in the constitutive expression of cut 2 that causes production of low levels of cutin monomers that strongly induce cut 1 using CTF1 as the transcription factor.