NDLI: Nod factors activate both heterotrimeric and monomeric G-proteins in Vigna unguiculata (L.) Walp

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Nod factors activate both heterotrimeric and monomeric G-proteins in Vigna unguiculata (L.) Walp

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Planta : Volume 216, Issue 3, January 2003

Planta : Volume 216, Issue 2, December 2002

Planta : Volume 216, Issue 1, November 2002

Nod factors activate both heterotrimeric and monomeric G-proteins in Vigna unguiculata (L.) Walp

Content Provider	Springer Nature Link
Author	Kelly, Marilyn N. Irving, Helen R.
Copyright Year	2003
Abstract	Nod factors are lipo-chito-oligosaccharides secreted by rhizobia that initiate many responses in the root hairs of the legume hosts, culminating in deformed hairs. The heterotrimeric G-protein agonists mastoparan, Mas7, melittin, compound 48/80 and cholera toxin provoke root hair deformation, whereas the heterotrimeric G-protein antagonist pertussis toxin inhibits mastoparan and Nod factor NodNGR[S]- (from Rhizobium sp. NGR234) induced root hair deformation. Another heterotrimeric G-protein antagonist, isotetrandrine, only inhibited root hair deformation provoked by mastoparan and melittin. These results support the notion that G-proteins are implicated in Nod factor signalling. To study the role of G-proteins at a biochemical level, we examined the GTP-binding profiles of root microsomal membrane fractions isolated from the nodulation competent zone of Vigna unguiculata (L.) Walp. GTP competitively bound to the microsomal membrane fractions labelled with [35S]GTPγS, yielding a two-site displacement curve with displacement constants (K i) of 0.58 µM and 0.16 mM. Competition with either ATP or GDP revealed a one-site displacement curve with K i of 4.4 and 29 µM, respectively, whereas ADP and UTP were ineffective competitors. The GTP-binding profiles of microsomal membrane fractions isolated from roots pretreated with either NodNGR[S] or the four-sugar, N-N'-N"-N'"-tetracetylchitotetraose (TACT) backbone of Nod factors were significantly altered compared with control microsomal fractions. To identify candidate proteins, membrane proteins were separated by SDS-PAGE and electrotransferred to nitrocellulose. GTP overlay experiments revealed that membrane fractions isolated from roots pretreated with NodNGR[S] or TACT contained two proteins (28 kDa and 25 kDa) with a higher affinity for GTPγS than control membrane fractions. Western analysis demonstrated that membranes from the pretreated roots contained more of another protein (~55 kDa) recognised by Gαcommon antisera. These results provide pharmacological and biochemical evidence supporting the contention that G-proteins are involved in Nod factor signalling and, importantly, implicate monomeric G-proteins in this process.
Starting Page	674
Ending Page	685
Page Count	12
File Format	PDF
ISSN	00320935
Journal	Planta
Volume Number	216
Issue Number	4
e-ISSN	14322048
Language	English
Publisher	Springer-Verlag
Publisher Date	2002-09-26
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Content Type	Text
Resource Type	Article
Subject	Genetics Plant Science

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Nod factors activate both heterotrimeric and monomeric G-proteins in Vigna unguiculata (L.) Walp