Inhibition of Bemisia tabaci vectored, GroEL mediated transmission of tomato leaf curl New Delhi virus by garlic leaf lectin (Allium sativum leaf agglutinin).

GroEL or symbionin synthesized by the endosymbionts of whitefly (Bemisia tabaci)/ aphids play a cardinal role in the persistent, circulative transmission of plant viruses by binding to viral coat protein/ read-through protein. Allium sativum leaf agglutinin (ASAL), a Galanthus nivalis agglutinin (GNA)- related mannose-binding lectin from garlic leaf has been reported as a potent controlling agent against hemipteran insects including whitefly and aphids. GroEL related chaperonin- symbionin was previously identified as a receptor of ASAL by the present group in the brush border membrane vesicle (BBMV) of mustard aphid. In the present study similar GroEL receptor of ASAL has been identified through LC-MS/MS in the BBMV of B. tabaci which serves as a vector for several plant viruses including tomato leaf curl New Delhi virus (ToLCNDV). Ligand blot analysis of ASAL-fed B. tabaci showed that when GroEL is pre-occupied by ASAL, it completely blocks its further binding to ToLCNDV coat protein (ToLCNDV-CP). Prior feeding of ASAL hindered the co-localization of ToLCNDV-CP and GroEL in the midgut of B. tabaci. Immunoprecipitation followed by western blot with ASAL-fed B. tabaci yielded similar result. Moreover, ASAL feeding inhibited viral transmission by B. tabaci. Together, these results confirmed that the interaction of ASAL with GroEL interferes with the binding of ToLCNDV-CP and inhibits further B. tabaci mediated viral transmission.

Thromboxane A2 receptor and MaxiK-channel intimate interaction supports channel trans-inhibition independent of G-protein activation.

Large conductance voltage- and calcium-activated potassium channels (MaxiK, BK(Ca)) are well known for sustaining cerebral and coronary arterial tone and for their linkage to vasodilator ß-adrenergic receptors. However, how MaxiK channels are linked to counterbalancing vasoconstrictor receptors is unknown. Here, we show that vasopressive thromboxane A2 receptors (TP) can intimately couple with and inhibit MaxiK channels. Activation of the receptor with its agonist trans-inhibits MaxiK independently of G-protein activation. This unconventional mechanism is supported by independent lines of evidence: (i) inhibition of MaxiK current by thromboxane A2 mimetic, U46619, occurs even when G-protein activity is suppressed; (ii) MaxiK and TP physically associate and display a high degree of proximity; and (iii) Förster resonance energy transfer occurs between fluorescently labeled MaxiK and TP, supporting a direct interaction. The molecular mechanism of MaxiK-TP intimate interaction involves the receptor's first intracellular loop and C terminus, and it entails the voltage-sensing conduction cassette of MaxiK channel. Further, physiological evidence of MaxiK-TP physical interaction is given in human coronaries and rat aorta, and by confirming TP role (with antagonist SQ29,548) in the U46619-induced MaxiK inhibition in human coronaries. We propose that vasoconstrictor TP receptor and MaxiK-channel direct interaction facilitates G-protein-independent TP to MaxiK trans-inhibition, which would promote vasoconstriction.