Redundancy in periplasmic binding protein-dependent transport systems for trehalose, sucrose, and maltose in Sinorhizobium meliloti.
We have identified a cluster of six genes involved in trehalose transport and utilization (thu) in Sinorhizobium meliloti. Four of these genes, thuE, -F, -G, and -K, were found to encode components of a binding protein-dependent trehalose/maltose/sucrose ABC transporter. Their deduced gene products comprise a trehalose/maltose-binding protein (ThuE), two integral membrane proteins (ThuF and ThuG), and an ATP-binding protein (ThuK). In addition, a putative regulatory protein (ThuR) was found divergently transcribed from the thuEFGK operon. When the thuE locus was inactivated by gene replacement, the resulting S. meliloti strain was impaired in its ability to grow on trehalose, and a significant retardation in growth was seen on maltose as well. The wild type and the thuE mutant were indistinguishable for growth on glucose and sucrose. This suggested a possible overlap in function of the thuEFGK operon with the aglEFGAK operon, which was identified as a binding protein-dependent ATP-binding transport system for sucrose, maltose, and trehalose. The K(m)s for trehalose transport were 8 +/- 1 nM and 55 +/- 5 nM in the uninduced and induced cultures, respectively. Transport and growth experiments using mutants impaired in either or both of these transport systems show that these systems form the major transport systems for trehalose, maltose, and sucrose. By using a thuE'-lacZ fusion, we show that thuE is induced only by trehalose and not by cellobiose, glucose, maltopentaose, maltose, mannitol, or sucrose, suggesting that the thuEFGK system is primarily targeted toward trehalose. The aglEFGAK operon, on the other hand, is induced primarily by sucrose and to a lesser extent by trehalose. Tests for root colonization, nodulation, and nitrogen fixation suggest that uptake of disaccharides can be critical for colonization of alfalfa roots but is not important for nodulation and nitrogen fixation per se.
Jensen
John Beck
JB
Department of Biology, Faculty of Science, University of Tromsø, N-9037 Tromsø, Norway.
Peters
N Kent
NK
Bhuvaneswari
T V
TV
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D001692:Biological Transport / Q000235:genetics*; D003470:Culture Media; D005798:Genes, Bacterial; D005947:Glucose; D008320:Maltose / Q000378:metabolism*; D000455:Medicago sativa / Q000382:microbiology; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009876:Operon; D016962:Sinorhizobium meliloti / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D013395:Sucrose / Q000378:metabolism*; D014199:Trehalose / Q000378:metabolism*
D018528:ATP-Binding Cassette Transporters; D003470:Culture Media; D013395:Sucrose; D008320:Maltose; D014199:Trehalose; D005947:Glucose