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Characterization of pco-1, a newly identified gene which regulates purine catabolism in Neurospora

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Abstract

A new gene of Neurospora crassa, designated pco-1, was characterized and shown to regulate the expression of several genes which encode enzymes required for the catabolism of purines. Unlike the wild type, a pco-1 mutant created by repeat-induced point mutation cannot utilize purines as a nitrogen source. The PCO1 protein contains a Zn(II)2Cys6 binuclear cluster motif near its N-terminus, followed by a putative coiled-coil motif. A chemical crosslinking experiment demonstrated that PCO1 forms homodimers. PCO1 binds to CGG-N6-CCG elements located in the upstream promoter region of four genes encoding purine catabolic enzymes. Northern blot analysis demonstrated that a functional PCO1 protein is required for induction of xdh, which encodes xanthine dehydrogenase. Moreover, PCO1 was required for induction of three different purine catabolic enzymes. Two glutamine-rich domains occur in the C-terminal region of PCO1 and at least one of the glutamine-rich regions is required for PCO1 function, suggesting that they might play a role in transcriptional activation. The PCO1 protein does not interact with the global-acting NIT2 protein or the negative-acting NMR protein that functions in nitrogen catabolite repression. Induction of the xdh gene and synthesis of xanthine dehydrogenase is completely dependent upon PCO1, but does not require the global-acting NIT2 protein, suggesting that it is controlled by a novel regulatory mechanism.

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  1. Department of Biochemistry, The Ohio State University, Columbus, OH 43210, USA

    T. D. Liu & G. A. Marzluf

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  1. T. D. Liu
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  2. G. A. Marzluf
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Communicated by U. Kück

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Liu, T.D., Marzluf, G.A. Characterization of pco-1, a newly identified gene which regulates purine catabolism in Neurospora. Curr Genet 46, 213–227 (2004). https://doi.org/10.1007/s00294-004-0530-8

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