Characterization of the amyR gene encoding a transcriptional activator for the amylase genes in Aspergillus nidulans

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

“…The genetically identified malA gene encodes AmyR, and the malA1 allele carries a loss of function mutation (H478 to L) in AmyR. 27) The malA1 mutant has little amylolytic activity, resulting in the formation of diffuse colonies with extremely reduced conidiation on maltose or starch as the sole carbon source. We describe malA1 as amyR1/malA1 and its product as AmyR H478L in this manuscript to avoid confusion.…”

“…MA1 is a y To whom correspondence should be addressed. Fax: +81-52-789-4087; E-mail: koba@agr.nagoya-u.ac.jp progeny of the cross between ABPU1 (biA1 pyrG89; wA3; argB2; pyroA4) and FGSC A57 (yA2; pyroA4; amyR1/malA1), 27) and SA1 was made by a cross between ÁamyR103 and DAT1 in this study. The A. nidulans strains were grown at 37 C in standard minimal medium 31) supplemented with appropriate requirements unless stated otherwise.…”

“…This plasmid carried the amyRencoding 4.6-kb EcoR I fragment on pUC119 in the orientation opposite to the previously described pAN8. 27) The alcAp-GFP-AmyR fusion gene was constructed in two steps, as follows: First, the Hind III and Nco I digested alcA promoter fragment (0.5-kb) was placed between the Hind III and Nco I sites located immediately in front of the GFP gene on pCaMV35S-sGFP-nos3 0 . Then the BsrG I and EcoR I digested amyR fragment (2.5-kb) was introduced between the BsrG I and EcoR I sites of the resulting plasmid so that the amyR gene was fused in-frame to the C-terminus of the GFP gene.…”

“…pAR-Zn1 to Zn4 carry the genes for AmyR 1{224 , AmyR 1{411 , AmyR 1{514 , and AmyR 1{588 respectively. For expression of GFP-AmyR H478L , which carries the H478L mutation (amyR1/malA1), the 1.3-kb DNA fragment containing the mutation site was amplified with M13 and ABIamyseq4 primers using pBM1, which carries the 3.8-kb Hind III-EcoR I fragment encoding the amyR gene of A. nidulans MA1, 27) as the template. The amplified fragment was digested with Xho I and EcoR I and replaced with the corresponding fragment on pAGAR-F, resulting in pAGAR-F H478L .…”

“…[22][23][24] The transcriptional activator essential for isomaltose-triggered amylase induction is AmyR, a Zn(II) 2 Cys 6 transcriptional activator. [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.…”