Methods in Electron Microscope Autoradiography

Dashek, William V.

doi:10.1007/978-1-59259-232-6_17

Cited by 1 publication

(1 citation statement)

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“…To evaluate the relative distribution of the tracer in the nucleus and cytoplasm, we performed electron microscopic autoradiography analyses as previously described (Farquhar et al, 1978;Krueger et al, 1985;Dashek, 2000). The callus cells were incubated with 0.4 mM 3 H-JA (10 mCi/nmol, American Radiolabeled Chemicals) in 100 mM potassium phosphate buffer (pH 5.8) for 2 h and fixed in 4% glutaraldehyde in 100 mM potassium phosphate buffer for 3 h. The samples were then postfixed in 1% osmium tetroxide for 2 h. After three rinses in potassium phosphate buffer, the cells were rapidly dehydrated in single changes of 30%, 50%, 70%, 80%, 90%, and 100% acetone.…”

Section: Electron Microscopic Autoradiographymentioning

confidence: 99%

Transporter-Mediated Nuclear Entry of Jasmonoyl-Isoleucine Is Essential for Jasmonate Signaling

Zheng

et al. 2017

Molecular Plant

111

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To control gene expression by directly responding to hormone concentrations, both animal and plant cells have exploited comparable mechanisms to sense small-molecule hormones in nucleus. Whether nuclear entry of these hormones is actively transported or passively diffused, as conventionally postulated, through the nuclear pore complex, remains enigmatic. Here, we identified and characterized a jasmonate transporter in Arabidopsis thaliana, AtJAT1/AtABCG16, which exhibits an unexpected dual localization at the nuclear envelope and plasma membrane. We show that AtJAT1/AtABCG16 controls the cytoplasmic and nuclear partition of jasmonate phytohormones by mediating both cellular efflux of jasmonic acid (JA) and nuclear influx of jasmonoyl-isoleucine (JA-Ile), and is essential for maintaining a critical nuclear JA-Ile concentration to activate JA signaling. These results illustrate that transporter-mediated nuclear entry of small hormone molecules is a new mechanism to regulate nuclear hormone signaling. Our findings provide an avenue to develop pharmaceutical agents targeting the nuclear entry of small molecules.

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Section: Electron Microscopic Autoradiographymentioning

confidence: 99%