Expression cloning in Fe2+ transport defective yeast of a novel maize MYC transcription factor

被引:14
作者
Loulergue, C
Lebrun, M
Briat, JF
机构
[1] Univ Montpellier 2, Inst Natl Rech Agron, CNRS, URA 2133, F-34060 Montpellier, France
[2] Ecole Natl Super Agron Montpellier, F-34060 Montpellier, France
[3] Univ Montpellier 2, F-34095 Montpellier, France
关键词
iron; roots; plants; R proteins;
D O I
10.1016/S0378-1119(98)00531-9
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
A complementation approach of the yeast fet3fet4 mutant strain, defective in both low- and high-affinity iron transport, was initiated as an attempt to characterize the Fe(III)-mugineic acid (MA) transporter from grasses. A maize cDNA encoding a novel MYC transcription factor, named 7E, was cloned by screening an iron-deficient maize root cDNA expression library on a minimum media containing Fe(III)-deoxyMA as a unique iron source. 7E expression restored growth specifically to the fet3 fet4 mutant strain. It did not affect growth rate of a trk1 trk2 potassium transport defective yeast strain or parental W303 strain growth rate, No Fe-55 uptake increase was observed in 7E transformed fet3 fet4 yeast during short-term kinetics. However, the iron accumulation in these cells was 1.3-fold higher than in untransformed cells after a 24-h period. The 7E protein contained 694 amino acids and had a predicted molecular mass of 74.2 kDa. It had 44% identity with the RAP-1 protein, a 67.9-kDa MYC-like protein from Ambidopsis thaliana which binds the G-box sequence via a basic region helix-loop-helix (bHLH), without requiring heterodimerization with MYB proteins. Phylogenic comparisons revealed that the maize 7E protein was related to the Arabidopsis thaliana RAP-1 protein and to the Phaseolus vulgaris PGI. This similarity was particularly evident for the bHLH domain, which was 95% identical between maize 7E and Arabidopsis thaliana RAP-1. 7E, RAP-I and PG-I proteins revealed a plant MYC-like sub-family that was more related to the maize repressor-like IN1 than to maize R proteins. 7E mRNA was detected in both roots and leaves by the Northern analysis. The amount of 7E mRNA increased, in response to iron starvation, by 20 and 40% in roots and leaves, respectively. The relationship between iron metabolism and myc expression in animal cells is discussed. (C) 1998 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:47 / 57
页数:11
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