In Vivo and in Vitro Analyses of the AmyR Binding Site of the Aspergillus nidulans agdA Promoter; Requirement of the CGG Direct Repeat for Induction and High Affinity Binding of AmyR

Inducer-Dependent Nuclear Localization of a Zn(II)2Cys6Transcriptional Activator, AmyR, inAspergillus nidulans

“…Rabbit antiserum against MalEAmyR 1{411 was prepared with purified MalE-AmyR 1{411 . 28) Rabbit anti-MalE-AmyR 1{411 antibody was purified from the antiserum using the MalE-AmyR 1{411 column, which was made from HiTrapÔ NHSactivated HP (Amersham Pharmacia Biotech, Uppsala, Sweden) and purified MalE-AmyR 1{411 following the manufacturer's instructions. The antibody was eluted from the column with 0.1 M glycine-HCl (pH 2.7) containing 0.5 M NaCl.…”

“…[25][26][27] AmyR binds to the direct repeat of CGG triplets separated by 8 bases in the various amylase promoter regions or the CGGN 8 AGG site of the Taka-amylase A promoter. 25,[28][29][30] In this study, subcellular localization studies and C-terminal truncation analysis of AmyR were carried out to obtain insight into the mechanisms of AmyR mediated induction. We found that nuclear accumulation of the GFP-AmyR fusion protein is triggered by the -amylase inducer isomaltose.…”

Mode of AmyR Binding to the CGGN8AGG Sequence in theAspergillus oryzaetaaG2Promoter

“…Mutational analysis of the 5 0 -CGGN 8 CGG-3 0 -type AmyR binding site of the A. nidulans -glucosidase A (agdA) promoter indicates that AmyR can bind to a single CGG triplet as a monomer but requires the CGG direct repeat for high affinity binding and transcriptional activation. 16) The question arises how the 5 0 -CGGAAATTTAA-3 0 sequence carrying only the single CGG triplet can function as an upstream activating sequence responsible for induction. In this study, site-directed mutational analysis of the AmyR binding site on the taaG2 promoter indicated that the AGG triplet neighboring the 3 0 -end of 5 0 -CGGAAATTTAA-3 0 is also required for binding of two AmyR molecules as well as induction of the taaG2 gene.…”

Molecular Analysis of AsamyR Gene Encoding Transcriptional Factor for Amylolytic Gene from Shoyu Koji Mold, Aspergillus sojae KBN1340

“…The DNA binding domain of AsAmyR shares a completely identical sequence with those of A. nidulans and A. oryzae (Fig. 1B), implying that AsAmyR recognizes and binds the same sequence, CGGN 8 (C/ A)GG, as these AmyRs (Petersen et al, 1999, Gomi et al, 2000, Tani et al, 2001a, Tani et al, 2001b, Itoh et al, 2004, Nakamura et al, 2006. Furthermore, the other four domains of AsAmyR showed significant sequence identity with the A. on the conserved domain structure between A. nidulans AmyR and Saccharomyces cerevisiae Mal63p, Makita et al presumed that the inducer-dependent nuclear localization of AmyR in A. nidulans is controlled by Hsp90/Hsp70-based chaperone machinery.…”

“…AmyR is a specific transcriptional activator with Zn(II) 2 Cys 6 binuclear cluster DNA-binding motif. AmyR binds the CGGN 8 AGG sequence at approximately -200 in the taaG2 gene promoter which is required for inducible expression by starch or maltose (Petersen et al, 1999, Gomi et al, 2000, Tani et al, 2001a, Itoh et al, 2004.…”