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THE PLANT CELL, Vol 2, Issue 9 925-939, Copyright © 1990 by American Society of Plant Biologists
Nuclear Factors Interact with Conserved A/T-Rich Elements Upstream of a Nodule-Enhanced Glutamine Synthetase Gene from French Bean
B. G. Forde, J. Freeman, J. E. Oliver and M. Pineda
Biochemistry and Physiology Department, AFRC Institute of Arable Crops Research, Rothamsted Experimental Station, Harpenden AL5 2JQ, United Kingdom
The gln-[gamma] gene, encoding the [gamma] subunit of glutamine synthetase
in French bean (Phaseolus vulgaris), is strongly induced during nodule
development. We have determined the nucleotide sequence of a 1.3-kilobase
region at its 5[prime] end and have identified several sequences common to
the promoter regions of late nodulin genes from other legume species. The
5[prime]-flanking region was analyzed for sequence-specific interactions
with nuclear factors from French bean. A factor from nodules (PNF-1) was
identified that binds to multiple sites between -860 and -154, and a
related but distinct factor (PRF-1) was detected in extracts from
uninfected roots. PNF-1 and PRF-1 bound strongly to a synthetic
oligonucleotide containing the sequence of an A/T-rich 21-base pair
imperfect repeat found at positions -516 and -466. The same factors also
had a high affinity for a protein binding site from a soybean leghemoglobin
gene and appeared to be closely related to the soybean nodule factor NAT2,
which binds to A/T-rich sequences in the lbc3 and nodulin 23 genes
[Jacobsen et al. (1990). Plant Cell 2, 85-94]. Comparison of NAT2/PNF-1
binding sites from a variety of nodulin genes revealed the conservation of
the short consensus core motif TATTTWAT, and evidence was obtained that
this sequence is important for protein recognition. Cross-recognition by
PNF-1 of a protein binding site in a soybean seed protein gene points to
the existence of a ubiquitous family of factors with related binding
affinities. Our data suggest that PNF-1 and PRF-1 belong to an
evolutionarily conserved group of nuclear factors that interact with
specific A/T-rich sequences in a diverse set of plant genes. We consider
the possible role of these factors in coregulating the expression of
gln-[gamma] and other late nodulin genes.
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