Investigating the DNA Binding Properties of the Initiator Binding Protein 2 (IBP2) in Maize (Zea Mays)
The maize (Zea mays L) initiator-binding protein2 (IBP2) gene is known to encode a protein, IBP2, with promoter-binding activity. IBP2 also resembles telomere DNA-binding proteins from other plant species. Therefore, we investigated whether the IBP2 protein, a presumed transcription factor, may actu...
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ndltd-fsu.edu-oai-fsu.digital.flvc.org-fsu_1805212020-06-09T03:09:22Z Investigating the DNA Binding Properties of the Initiator Binding Protein 2 (IBP2) in Maize (Zea Mays) Moore, Joel M. (authoraut) Bass, Hank W. (professor directing thesis) McGinnis, Karen M. (committee member) Tang, Hengli (committee member) Department of Biological Science (degree granting department) Florida State University (degree granting institution) Text text Florida State University Florida State University English eng 1 online resource computer application/pdf The maize (Zea mays L) initiator-binding protein2 (IBP2) gene is known to encode a protein, IBP2, with promoter-binding activity. IBP2 also resembles telomere DNA-binding proteins from other plant species. Therefore, we investigated whether the IBP2 protein, a presumed transcription factor, may actually have telomere DNA-binding activity. Using Electrophoretic Mobility Shift Assay (EMSA) we found that a C-terminal sublcone of the IBP2 protein, rIBP2 Myb-Q, binds telomeric repeat DNA in vitro. We also found that rIBP2 Myb-Q binds strongly to double stranded telomere repeat sequences with three or four tandem repeats, but only weakly to sequences with two tandem repeats. Furthermore, we found no evidence of binding with a single repeat (5' TTTAGGG 3') sequence. We also found that single point changes in (T1:A and G6:T) in the three-repeat sequence diminished, but did not entirely abolish, binding activity. Additionally, we found that the rIBP2 Myb-Q did bind weakly to the initiator sequence of the Shrunken1 promoter. From these experiments, we conclude that IBP2 is likely to have telomeric functions in vivo, and possibly play a role in both gene transcription and telomere functions. A Thesis Submitted to the Department of Biological Science in Partial Fulfillment of the Requirements for the Degree of Master of Science. Fall Semester, 2009. November 2, 2009. Shelterin, Telomere, Maize, Binding, Protein, Plant Includes bibliographical references. Hank W. Bass, Professor Directing Thesis; Karen M. McGinnis, Committee Member; Hengli Tang, Committee Member. Biology FSU_migr_etd-2306 http://purl.flvc.org/fsu/fd/FSU_migr_etd-2306 This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). The copyright in theses and dissertations completed at Florida State University is held by the students who author them. http://diginole.lib.fsu.edu/islandora/object/fsu%3A180521/datastream/TN/view/Investigating%20the%20DNA%20Binding%20Properties%20of%20the%20Initiator%20Binding%20Protein%202%20%28IBP2%29%20in%20Maize%20%28Zea%20Mays%29.jpg |
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Biology Investigating the DNA Binding Properties of the Initiator Binding Protein 2 (IBP2) in Maize (Zea Mays) |
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The maize (Zea mays L) initiator-binding protein2 (IBP2) gene is known to encode a protein, IBP2, with promoter-binding activity. IBP2 also resembles telomere DNA-binding proteins from other plant species. Therefore, we investigated whether the IBP2 protein, a presumed transcription factor, may actually have telomere DNA-binding activity. Using Electrophoretic Mobility Shift Assay (EMSA) we found that a C-terminal sublcone of the IBP2 protein, rIBP2 Myb-Q, binds telomeric repeat DNA in vitro. We also found that rIBP2 Myb-Q binds strongly to double stranded telomere repeat sequences with three or four tandem repeats, but only weakly to sequences with two tandem repeats. Furthermore, we found no evidence of binding with a single repeat (5' TTTAGGG 3') sequence. We also found that single point changes in (T1:A and G6:T) in the three-repeat sequence diminished, but did not entirely abolish, binding activity. Additionally, we found that the rIBP2 Myb-Q did bind weakly to the initiator sequence of the Shrunken1 promoter. From these experiments, we conclude that IBP2 is likely to have telomeric functions in vivo, and possibly play a role in both gene transcription and telomere functions. === A Thesis Submitted to the Department of Biological Science in Partial Fulfillment of the Requirements for the Degree of Master of Science. === Fall Semester, 2009. === November 2, 2009. === Shelterin, Telomere, Maize, Binding, Protein, Plant === Includes bibliographical references. === Hank W. Bass, Professor Directing Thesis; Karen M. McGinnis, Committee Member; Hengli Tang, Committee Member. |
author2 |
Moore, Joel M. (authoraut) |
author_facet |
Moore, Joel M. (authoraut) |
title |
Investigating the DNA Binding Properties of the Initiator Binding Protein 2 (IBP2) in Maize (Zea Mays) |
title_short |
Investigating the DNA Binding Properties of the Initiator Binding Protein 2 (IBP2) in Maize (Zea Mays) |
title_full |
Investigating the DNA Binding Properties of the Initiator Binding Protein 2 (IBP2) in Maize (Zea Mays) |
title_fullStr |
Investigating the DNA Binding Properties of the Initiator Binding Protein 2 (IBP2) in Maize (Zea Mays) |
title_full_unstemmed |
Investigating the DNA Binding Properties of the Initiator Binding Protein 2 (IBP2) in Maize (Zea Mays) |
title_sort |
investigating the dna binding properties of the initiator binding protein 2 (ibp2) in maize (zea mays) |
publisher |
Florida State University |
url |
http://purl.flvc.org/fsu/fd/FSU_migr_etd-2306 |
_version_ |
1719318345774268416 |