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Cloning and molecular analyses of hop transcription factors

(2009) Acta Horticulturae. 848. p.41-48
Author
Organization
Abstract
Expression of hop chs_H1 and other hop chalcone synthase (CHS)-like enzymes was assayed in developing pollen tissue. Both, chs_H1 and valerophenone synthase (vps) are highly expressed in pollen suggesting the existence of common specific regulatory element(s). While vps is highly specific for female and male floral organs, the oligofamily of chs_H1 is expressed in both generative and somatic tissues, suggesting a more universal promoter. To identify novel transcription factors involved in the regulation of chs_H1 genes in lupulin glands, cone-specific ESTs showing homology to bZIP and bHLH were utilized to screen hop cDNA and genomic libraries. Additionally, inverse PCR was used to amplify full length cDNAs of members of two bZIP oligofamilies from hop. Transcription factors H/bZIP1A and H/bZIP2 have predicted molecular masses of 27.4 and 34.2 kDa, respectively. While H/bZIP1A is rather neutral (pI 6.42), H/bZIP2 is strongly basic (pI 8.51). A truncated variant of H/bZIP1 (H/bZIP1B) that is strongly basic but lacks the zipper domain was also cloned from hop. Additionally, the first bHLH transcription factor was cloned from hop. H/bHLH1 is a protein of 33.6 kDa with a predicted pI of 6.08. Based on semiquantitative RT PCR results, this TF is expressed specifically in hop floral and cone tissue. The combinatorial action of H/bZIP2 and other hop TFs that have been cloned, especially H/Myb1 and 3 were analyzed in P. hybrida and N. benthamiana transient expression systems. TF H/bZIP2, as well as both Myb TFs, were found as activators of chs_H1 promoter. In the combinatorial system, H/bZIP2 was found to modulate expression of H/Myb1 and H/Myb3 Trs from hop.
Keywords
REGULATORY FACTOR, BIOSYNTHESIS, CHALCONE SYNTHASE HOMOLOGS, HUMULUS-LUPULUS L., N. benthamiana, lupulin, cis-elements, hop metabolome, P. hybrida, transcription regulators, H. lupulus, real-time PCR, pollen development, Agrobacterium leaf infiltration, ACTIVATION, EXPRESSION, EVOLUTION, SEQUENCE, GENE, DNA, A. thaliana

Citation

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Chicago
Matoušek, Jaroslav, Thomas Kocábek, Josef Škopek, L Orctová, J Stehlik, Zoltan Füssy, Josef Patzak, et al. 2009. “Cloning and Molecular Analyses of Hop Transcription Factors.” In Acta Horticulturae, ed. Denis De Keukeleire and Kim E Hummer, 848:41–48. Leuven, Belgium: International Society for Horticultural Science (ISHS).
APA
Matoušek, J., Kocábek, T., Škopek, J., Orctová, L., Stehlik, J., Füssy, Z., Patzak, J., et al. (2009). Cloning and molecular analyses of hop transcription factors. In D. De Keukeleire & K. E. Hummer (Eds.), Acta Horticulturae (Vol. 848, pp. 41–48). Presented at the 2nd International Humulus Symposium, Leuven, Belgium: International Society for Horticultural Science (ISHS).
Vancouver
1.
Matoušek J, Kocábek T, Škopek J, Orctová L, Stehlik J, Füssy Z, et al. Cloning and molecular analyses of hop transcription factors. In: De Keukeleire D, Hummer KE, editors. Acta Horticulturae. Leuven, Belgium: International Society for Horticultural Science (ISHS); 2009. p. 41–8.
MLA
Matoušek, Jaroslav, Thomas Kocábek, Josef Škopek, et al. “Cloning and Molecular Analyses of Hop Transcription Factors.” Acta Horticulturae. Ed. Denis De Keukeleire & Kim E Hummer. Vol. 848. Leuven, Belgium: International Society for Horticultural Science (ISHS), 2009. 41–48. Print.
@inproceedings{872777,
  abstract     = {Expression of hop chs\_H1 and other hop chalcone synthase (CHS)-like enzymes was assayed in developing pollen tissue. Both, chs\_H1 and valerophenone synthase (vps) are highly expressed in pollen suggesting the existence of common specific regulatory element(s). While vps is highly specific for female and male floral organs, the oligofamily of chs\_H1 is expressed in both generative and somatic tissues, suggesting a more universal promoter. To identify novel transcription factors involved in the regulation of chs\_H1 genes in lupulin glands, cone-specific ESTs showing homology to bZIP and bHLH were utilized to screen hop cDNA and genomic libraries. Additionally, inverse PCR was used to amplify full length cDNAs of members of two bZIP oligofamilies from hop. Transcription factors H/bZIP1A and H/bZIP2 have predicted molecular masses of 27.4 and 34.2 kDa, respectively. While H/bZIP1A is rather neutral (pI 6.42), H/bZIP2 is strongly basic (pI 8.51). A truncated variant of H/bZIP1 (H/bZIP1B) that is strongly basic but lacks the zipper domain was also cloned from hop. Additionally, the first bHLH transcription factor was cloned from hop. H/bHLH1 is a protein of 33.6 kDa with a predicted pI of 6.08. Based on semiquantitative RT PCR results, this TF is expressed specifically in hop floral and cone tissue. The combinatorial action of H/bZIP2 and other hop TFs that have been cloned, especially H/Myb1 and 3 were analyzed in P. hybrida and N. benthamiana transient expression systems. TF H/bZIP2, as well as both Myb TFs, were found as activators of chs\_H1 promoter. In the combinatorial system, H/bZIP2 was found to modulate expression of H/Myb1 and H/Myb3 Trs from hop.},
  author       = {Matou\v{s}ek, Jaroslav and Koc{\'a}bek, Thomas and \v{S}kopek, Josef and Orctov{\'a}, L and Stehlik, J and F{\"u}ssy, Zoltan and Patzak, Josef and Krofta, Karel  and Maloukh, Lina and Rold{\`a}n-Ruiz, Isabel and Heyerick, Arne and De Keukeleire, Denis},
  booktitle    = {Acta Horticulturae},
  editor       = {De Keukeleire, Denis and Hummer, Kim E},
  isbn         = {9789066057227},
  issn         = {0567-7572},
  keyword      = {REGULATORY FACTOR,BIOSYNTHESIS,CHALCONE SYNTHASE HOMOLOGS,HUMULUS-LUPULUS L.,N. benthamiana,lupulin,cis-elements,hop metabolome,P. hybrida,transcription regulators,H. lupulus,real-time PCR,pollen development,Agrobacterium leaf infiltration,ACTIVATION,EXPRESSION,EVOLUTION,SEQUENCE,GENE,DNA,A. thaliana},
  language     = {eng},
  location     = {Ghent, Belgium},
  pages        = {41--48},
  publisher    = {International Society for Horticultural Science (ISHS)},
  title        = {Cloning and molecular analyses of hop transcription factors},
  url          = {http://www.actahort.org/books/848/848\_3.htm},
  volume       = {848},
  year         = {2009},
}

Web of Science
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