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Do the tightly linked structural genes for nitrate and nitrite reductases in Aspergillus nidulans form an operon? Evidence from an insertional translocation which separates them

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Summary

Previous work (Rand and Arst, 1977) led to the proposal that the nis-5 mutation results in a new low activity promoter for niiA, the structural gene for nitrite reductase in Aspergillus nidulans. Expression of niiA via this promoter differs from expression of niiA via its normal promoter/initiator in that expression by the new promoter is not subject to nitrate induction or ammonium repression. nis-5 reduces but does not abolish niiA expression mediated by the normal promoter/initiator.

In this work we show that nis-5 is associated with and is probably identical to a non-reciprocal translocation in which a considerable portion of the centromere proximal region of the right arm of linkage group II is inserted into linkage group VIII between niiA and niaD, the tightly linked, probably contiguous structural genes for nitrate reductase. This implies that niiA, along with its normal promoter/initiator, has been fused to a promoter normally in linkage group IIR but as yet unidentified by its normal rôle. Further, it indicates that niiA is transcribed from the niaD-proximal side. As niiA and niaD are separated by a large number of unrelated genes in nis-5 strains, we can safely conclude that expression of niiA does not occur solely by synthesis of a messenger which carries a niaD as well as a niiA transcript. Clearly, niiA and niaD do not form an operon for which a di-(or poly-), cistronic messenger be the only transcript. This is consistent with other experimental evidence which shows that the synthesis of nitrate and nitrite reductases are not coordinately regulated. Nevertheless, all of these data would also be consistent with a model in which niiA and niaD form an operontype structure having overlapping transcripts, one being di-(or poly-) cistronic and including both niiA and niaD and another being monocistronic for niiA.

The reduced niiA expression mediated by the normal promoter/initiator in nis-5 strains could be a consequence of the functioning or positioning of the new linkage group II niiA promoter. An alternative, but not mutually exclusive, explanation would be that the insertional translocation prevents synthesis of a niiA niaD dicistronic transcript so that only that component of niiA expression which is due to a monocistronic niiA messenger can be induced by nitrate (and nitrite) in nis-5 strains. The apparently low activity of the new linkage group II promoter in comparison to the normal niiA promoter/initiator might betoken considerable efficiency of the latter rather than any particular lack of efficiency of the former.

In addition, this work has involved extensive new mapping in linkage group II, including both mitotic mapping of the centromere and meiotic mapping of previously unlocated markers. A series of crosses including genotype combinations both heterozygous and homozygous for nis-5 has been used to map the break-points and orientation of the translocation. As one break-point is closer to the centromere of linkage group II than the most centromere proximal identified gene on the same (i.e. right) arm we showed that the position of the centromere, as defined by mitotic recombination, has not been affected.

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Author notes

  1. Keith N. Rand

    Present address: Glaxo Laboratories Ltd, North Lonsdale Road, LA12 9DR, Ulverston, Cumbria, England

  2. Christopher R. Bailey

    Present address: Beecham Pharmaceuticals, Clarendon Road, BN14 8QH, Worthing, West Sussex, England

Authors and Affiliations

  1. Department of Genetics, University of Cambridge, Downing Street, CB2 3EH, Cambridge, England

    Herbert N. Arst Jr., Keith N. Rand & Christopher R. Bailey

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  1. Herbert N. Arst Jr.
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  2. Keith N. Rand
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Communicated by W. Gajewski

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Arst, H.N., Rand, K.N. & Bailey, C.R. Do the tightly linked structural genes for nitrate and nitrite reductases in Aspergillus nidulans form an operon? Evidence from an insertional translocation which separates them. Molec. Gen. Genet. 174, 89–100 (1979). https://doi.org/10.1007/BF00433309

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