Translation and assembly of CABYR coding region B in fibrous sheath and restriction of calcium binding to coding region A
Autor: | Soren Naaby-Hansen, Ken Klotz, Kula N. Jha, Young-Hwan Kim, Charles J. Flickinger, Erin M. Farris, Phillip L. Snow, John C. Herr, Arabinda Mandal, Geeta Vanage |
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Rok vydání: | 2005 |
Předmět: |
Gene isoform
Male DNA Complementary Population Sperm flagella Biology Antibodies law.invention 03 medical and health sciences 0302 clinical medicine Protein phosphorylation law Calcium-binding protein Capacitation Coding region Animals Humans Protein Isoforms Cloning Molecular education Spermatogenesis Molecular Biology 030304 developmental biology 0303 health sciences education.field_of_study 030219 obstetrics & reproductive medicine Polymorphism Genetic Base Sequence Alternative splicing Calcium-Binding Proteins CABYR Cell Biology Phosphoproteins Molecular biology Stop codon Recombinant Proteins Rats Alternative Splicing Microscopy Electron Polyclonal antibodies Protein Biosynthesis Sperm Tail biology.protein Recombinant DNA Codon Terminator Calcium Calcium binding Developmental Biology |
Zdroj: | Developmental Biology. 286(1):46-56 |
ISSN: | 0012-1606 |
DOI: | 10.1016/j.ydbio.2005.07.005 |
Popis: | CABYR is a highly polymorphic, sperm flagellar calcium-binding protein that is tyrosine as well as serine/threonine phosphorylated during capacitation. Six alternative splice variants of human CABYR (I–VI) have previously been identified, involving two coding regions, CR-A and CR-B, separated by an intervening stop codon. It is presently unknown if proteins encoded by the predicted coding region B of CABYR are translated during spermiogenesis, where they localize, or which CABYR isoforms bind calcium. Immunofluorescent and electron microscopic studies using polyclonal antibodies generated to the recombinant c-terminal 198 aa CABYR-B localized the isoforms containing CABYR-B to the ribs and longitudinal columns of the fibrous sheath in the principal piece of the flagellum. Antisera to recombinant CABYR-A and CABYR-B proteins recognized distinct populations of CABYR isoforms encoded by either CR-A alone and/or CR-B as well as a common population of CABYR isoforms. Only the recombinant CABYR-A and not the CABYR-B bound calcium in vitro, which is consistent with the hypothesis that CABYR-A is the only form that binds calcium in sperm. These observations confirmed that, despite the presence of the stop codon in CR-A, splice variants containing CR-B are expressed during spermiogenesis and assemble into the fibrous sheath of the principal piece; however, calcium binding occurs only to those CABYR isoforms containing CABYR-A. |
Databáze: | OpenAIRE |
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