Expression of a calmodulin methylation mutant affects the growth and development of transgenic tobacco plants

Transgenic plants were constructed that express two foreign calmodulins (VU-1 and VU-3 calmodulins) derived from a cloned synthetic calmodulin gene. VU-1 calmodulin, similar to endogenous plant calmodulin, possesses a lysine residue at position 115 and undergoes posttranslational methylation. VU-3 c...

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Published in:Proceedings of the National Academy of Sciences - PNAS Vol. 89; no. 17; pp. 8394 - 8398
Main Authors: Roberts, D.M. (University of Tennessee, Knoxville, TN), Besl, L, Oh, S.H, Masterson, R.V, Schell, J, Stacey, G
Format: Journal Article
Language:English
Published: Washington, DC National Acad Sciences 01-09-1992
National Academy of Sciences of the United States of America
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Abstract Transgenic plants were constructed that express two foreign calmodulins (VU-1 and VU-3 calmodulins) derived from a cloned synthetic calmodulin gene. VU-1 calmodulin, similar to endogenous plant calmodulin, possesses a lysine residue at position 115 and undergoes posttranslational methylation. VU-3 calmodulin is a site-directed mutant of VU-1 calmodulin that is identical in sequence except for the substitution of an arginine at position 115 and thus is incapable of methylation. Both calmodulin genes, under the control of the cauliflower mosaic virus 35S promoter, were expressed in transgenic tobacco. Foreign calmodulin protein accumulated in plant tissues to levels equivalent to that of the endogenous calmodulin. All transformed lines of VU-1 plants were indistinguishable from untransformed controls with respect to growth and development. However, all transformed lines of VU-3 plants were characterized by decreased stem internode growth, reduced seed production, and reduced seed and pollen viability. The data suggest that these phenotypes are the result of the expression of the calmodulin mutant rather than the position of transferred DNA insertion or the overall alteration of calmodulin levels. Analyses of the activity of the purified transgenic calmodulins suggest that calmodulin-dependent NAD kinase is among the potential targets that may have altered regulation in VU-3 transgenic plants
AbstractList Transgenic plants were constructed that express two foreign calmodulins (VU-1 and VU-3 calmodulins) derived from a cloned synthetic calmodulin gene. VU-1 calmodulin, similar to endogenous plant calmodulin, possesses a lysine residue at position 115 and undergoes posttranslational methylation. VU-3 calmodulin is a site-directed mutant of VU-1 calmodulin that is identical in sequence except for the substitution of an arginine at position 115 and thus is incapable of methylation. Both calmodulin genes, under the control of the cauliflower mosaic virus 35S promoter, were expressed in transgenic tobacco. Foreign calmodulin protein accumulated in plant tissues to levels equivalent to that of the endogenous calmodulin. All transformed lines of VU-1 plants were indistinguishable from untransformed controls with respect to growth and development. However, all transformed lines of VU-3 plants were characterized by decreased stem internode growth, reduced seed production, and reduced seed and pollen viability. The data suggest that these phenotypes are the result of the expression of the calmodulin mutant rather than the position of transferred DNA insertion or the overall alteration of calmodulin levels. Analyses of the activity of the purified transgenic calmodulins suggest that calmodulin-dependent NAD kinase is among the potential targets that may have altered regulation in VU-3 transgenic plants.
Transgenic plants were constructed that express two foreign calmodulins (VU-1 and VU-3 calmodulins) derived from a cloned synthetic calmodulin gene. VU-1 calmodulin, similar to endogenous plant calmodulin, possesses a lysine residue at position 115 and undergoes posttranslational methylation. VU-3 calmodulin is a site-directed mutant of VU-1 calmodulin that is identical in sequence except for the substitution of an arginine at position 115 and thus is incapable of methylation. Both calmodulin genes, under the control of the cauliflower mosaic virus 35S promoter, were expressed in transgenic tobacco. Foreign calmodulin protein accumulated in plant tissues to levels equivalent to that of the endogenous calmodulin. All transformed lines of VU-1 plants were indistinguishable from untransformed controls with respect to growth and development. However, all transformed lines of VU-3 plants were characterized by decreased stem internode growth, reduced seed production, and reduced seed and pollen viability. The data suggest that these phenotypes are the result of the expression of the calmodulin mutant rather than the position of transferred DNA insertion or the overall alteration of calmodulin levels. Analyses of the activity of the purified transgenic calmodulins suggest that calmodulin-dependent NAD kinase is among the potential targets that may have altered regulation in VU-3 transgenic plants
Transgenic plants were constructed that express two foreign calmodulins (VU-1 and VU-3 calmodulins) derived from a cloned synthetic calmodulin gene. VU-1 calmodulin, similar to endogenous plant calmodulin, possesses a lysine residue at position 115 and undergoes posttranslational methylation. VU-3 calmodulin is a site-directed mutant of VU-1 calmodulin that is identical in sequence except for the substitution of an arginine at position 115 and thus is incapable of methylation. Both calmodulin genes, under the control of the cauliflower mosaic virus 35S promoter, were expressed in transgenic tobacco. Foreign calmodulin protein accumulated in plant tissue to levels equivalent to that of the endogenous calmodulin. All transformed lines of VU-1 plants were indistinguishable from untransformed controls with respect to growth and development. All transformed lines of VU-3 plants were characterized by decreased stem internode growth, reduced seed production, and reduced seed and pollen viability. The data suggest that these phenotypes are the result of the expression of the calmodulin mutant rather than the position of transferred DNA insertion or the overall alteration of calmodulin levels.
The expression of a calmodulin methylation mutant was found to affect the growth and development of transgenic tobacco plants.
Author Oh, S.H
Schell, J
Roberts, D.M. (University of Tennessee, Knoxville, TN)
Masterson, R.V
Besl, L
Stacey, G
AuthorAffiliation Center for Legume Research, University of Tennessee, Knoxville 37996
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Issue 17
Keywords Calcium
Nicotiana tabacum
Structure function relationship
Binding protein
Dicotyledones
Angiospermae
Development
Spermatophyta
Transgenic plant
Solanaceae
Mutation
Methylation
Calmodulin
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Snippet Transgenic plants were constructed that express two foreign calmodulins (VU-1 and VU-3 calmodulins) derived from a cloned synthetic calmodulin gene. VU-1...
The expression of a calmodulin methylation mutant was found to affect the growth and development of transgenic tobacco plants.
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SubjectTerms Amino Acid Sequence
Biochemistry
Biological and medical sciences
CALCIO
CALCIUM
Calmodulin - genetics
Calmodulin - metabolism
Cell differentiation, maturation, development, hematopoiesis
Cell physiology
CRECIMIENTO
CROISSANCE
Enzyme Activation
ETAPAS DE DESARROLLO DE LA PLANTA
EXPRESION GENICA
EXPRESSION DES GENES
Fundamental and applied biological sciences. Psychology
GENETICA
GENETIQUE
METHYLATION
METILACION
Molecular and cellular biology
Molecular Sequence Data
MUTANT
MUTANTES
Mutation
Nicotiana - genetics
NICOTIANA TABACUM
Phosphotransferases (Alcohol Group Acceptor)
Phosphotransferases - metabolism
PLANTAS TRANSGENICAS
PLANTE TRANSGENIQUE
Plants, Genetically Modified - growth & development
Plants, Toxic
Protein Processing, Post-Translational
PROTEINAS
PROTEINE
Proteins
Reproduction
STADE DE DEVELOPPEMENT VEGETAL
Structure-Activity Relationship
Tobacco
Title Expression of a calmodulin methylation mutant affects the growth and development of transgenic tobacco plants
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