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Reversible heat-induced inactivation of chimeric β-glucuronidase in transgenic plants

AuthorsAlmoguera, Concepción ; Rojas, Anabel ; Jordano, Juan
KeywordsTransgenic tobacco
Nicotiana tabacum
Translational fusion
Transcriptional fusion
Helianthus annuus
Transgenic plants
Plant proteins
Heat stress
Issue Date9-Apr-2002
PublisherAmerican Society of Plant Biologists
CitationPlant Physiology 129(1): 333–341, 2002
AbstractWe compared the expression patterns in transgenic tobacco (Nicotiana tabacum) of two chimeric genes: a translational fusion to ß-glucuronidase (GUS) and a transcriptional fusion, both with the same promoter and 5'-flanking sequences of Ha hsp17.7 G4, a small heat shock protein (sHSP) gene from sunflower (Helianthus annuus). We found that immediately after heat shock, the induced expression from the two fusions in seedlings was similar, considering chimeric mRNA or GUS protein accumulation. Surprisingly, we discovered that the chimeric GUS protein encoded by the translational fusion was mostly inactive in such conditions. We also found that this inactivation was fully reversible. Thus, after returning to control temperature, the GUS activity was fully recovered without substantial changes in GUS protein accumulation. In contrast, we did not find differences in the in vitro heat inactivation of the respective GUS proteins. Insolubilization of the chimeric GUS protein correlated with its inactivation, as indicated by immunoprecipitation analyses. The inclusion in another chimeric gene of the 21 amino-terminal amino acids from a different sHSP lead to a comparable reversible inactivation. That effect not only illustrates unexpected post-translational problems, but may also point to sequences involved in interactions specific to sHSPs and in vivo heat stress conditions.
Publisher version (URL)http://www.plantphysiol.org/cgi/content/abstract/129/1/333
ISBN1532-2548 Online
Appears in Collections:(IRNAS) Artículos
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