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Journal of Bacteriology, March 1999, p. 1544-1554, Vol. 181, No. 5
Center for Legume
Research,1 Department of
Microbiology,2 Department of
Botany,3 and Department of Ecology and
Evolutionary Biology,5 The University of
Tennessee, Knoxville, Tennessee 37996, and Department of
Soil, Water and Climate, The University of Minnesota, St. Paul,
Minnesota 551084
Received 27 August 1998/Accepted 18 December 1998
Examination of nolA revealed that NolA can be uniquely
translated from three ATG start codons. Translation from the first ATG
(ATG1) predicts a protein (NolA1) having an N-terminal,
helix-turn-helix DNA-binding motif similar to the DNA-binding domains
of the MerR-type regulatory proteins. Translation from ATG2 and ATG3
would give the N-terminally truncated proteins NolA2 and
NolA3, respectively, lacking the DNA-binding domain.
Consistent with this, immunoblot analyses of Bradyrhizobium
japonicum extracts with a polyclonal antiserum to NolA revealed
three distinct polypeptides whose molecular weights were consistent
with translation of nolA from the three ATG initiation
sites. Site-directed mutagenesis was used to produce derivatives of
nolA in which ATG start sites were sequentially deleted.
Immunoblots revealed a corresponding absence of the polypeptide whose
ATG start site was removed. Translational fusions of the nolA mutants to a promoterless lacZ yielded
functional fusion proteins in both Escherichia coli and
B. japonicum. Expression of NolA is inducible upon addition
of extracts from 5-day-old etiolated soybean seedlings but is not
inducible by genistein, a known inducer of the B. japonicum
nod genes. The expression of both NolA2 and
NolA3 requires the presence of NolA1.
NolA1 or NolA3 is required for the
genotype-specific nodulation of soybean genotype PI 377578.
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
The Bradyrhizobium japonicum nolA Gene
Encodes Three Functionally Distinct Proteins
*
Corresponding author. Mailing address: Center for
Legume Research, Department of Microbiology, M409 Walters Life Science
Building, The University of Tennessee, Knoxville, TN 37996-0845. Phone: (423) 974-4041. Fax: (423) 974-4007. E-mail:
GSTACEY{at}utk.edu.
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