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THE PLANT CELL, Vol 2, Issue 1 85-94, Copyright © 1990 by American Society of Plant Biologists
HMG I-Like Proteins from Leaf and Nodule Nuclei Interact with Different AT Motifs in Soybean Nodulin Promoters
K. Jacobsen, N. B. Laursen, E. O. Jensen, A. Marcker, C. Poulsen and K. A. Marcker
Department of Molecular Biology and Plant Physiology, University of Aarhus, C.F. Mollers Alle 130, DK-8000 Aarhus C, Denmark
Three different nuclear factors recognizing short AT-rich DNA sequences
were identified in different organs of soybean. One factor (NAT2) was found
to be present in mature nodules, another factor (NAT1) was detected in
roots and nodules, and a third one (LAT1) was only observed in leaves. All
three factors recognized several DNA sequences in the promoter region of
the soybean nodulin N23 gene. Footprinting, deletion, and point mutation
analyses revealed different binding properties for all three factors and
further showed that even single base pair substitutions had a dramatic
effect on binding affinity. The LAT1 and NAT1 factors were released from
chromatin by extraction with a low-salt buffer and were soluble in 2%
trichloroacetic acid, implying a relationship to high-mobility group (HMG)
proteins. DNA binding studies further indicated a functional relationship
of these factors to the human HMG I protein. Purification of the LAT1
factor from leaf nuclei revealed the presence of two polypeptides with
molecular masses of 21 kilodaltons and 23 kilodaltons, respectively,
binding the same DNA sequence with equal affinity.
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