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A monoclonal antibody to lateral element proteins in synaptonemal complexes of Lilium longiflorum
L K Anderson1, S M Stack, R J Todd
1Department of Biology, Colorado State University, Fort Collins 80523.
Insights
Researchers identified synaptonemal complex (SC) proteins in lilies using a novel antibody. This antibody specifically targets lateral element proteins, suggesting a link between SC proteins and those in actively dividing cells.
Area of Science:
- Plant molecular biology
- Cell biology
- Genetics
Background:
- The synaptonemal complex (SC) is crucial for homologous chromosome pairing during meiosis.
- Identifying specific SC proteins is essential for understanding meiotic recombination and chromosome segregation.
- Lilium longiflorum (lily) is a model organism for studying meiosis due to its large chromosomes.
Purpose of the Study:
- To identify and characterize synaptonemal complex (SC) proteins in Lilium longiflorum.
- To develop specific antibodies for detecting SC components.
- To investigate the relationship between SC proteins and nuclear proteins in different cell types.
Main Methods:
- Generation of monoclonal antibodies by immunizing mice with isolated pachytene nuclei.
- Immunofluorescence microscopy to localize antibody binding on SCs.
- DNase digestion assays to differentiate SC proteins from surrounding nuclear components.
- Immunoblotting to identify protein targets of the antibody in various lily tissues.
Main Results:
- A specific monoclonal antibody (anti-LE) was generated, recognizing lateral element proteins of the SC.
- Anti-LE binding was enhanced on DNase-treated SCs, confirming specificity for lateral element proteins.
- Immunoblotting revealed that anti-LE recognizes proteins of various molecular weights (52,000–70,000 Da) in pachytene, tetrad, and root tip nuclei.
- Anti-LE did not bind to proteins from leaf nuclei, which lack actively dividing cells.
Conclusions:
- The study successfully identified a specific antibody against lily SC lateral element proteins.
- These findings suggest that some SC proteins are conserved and present in mitotically active cells.
- This research provides a valuable tool for further investigation into SC structure and function in plants.
Abstract:
To identify synaptonemal complex (SC) proteins in Lilium longiflorum (lily), monoclonal antibodies were generated using mice immunized with isolated pachytene nuclei. While most of the resulting monoclonal antibodies recognized nucleolar or chromatin proteins, one monoclonal antibody (anti-LE) was found that binds to lateral elements. Anti-LE bound more to lateral elements of SCs digested with DNase than to lateral elements that had not been digested with DNase. The opposite pattern of labeling was observed using monoclonal antibodies to lily chromatin and nucleolar proteins. These results indicate that anti-LE is specifically recognizing lateral element proteins and not chromatin or nucleolar proteins surrounding the lateral elements. On immunoblots, anti-LE binds to three pachytene nuclear proteins (Mr 60000, 66000 and 70000), two tetrad (early microspore) nuclear proteins (Mr 60000 and 70000), and two root tip nuclear proteins (Mr 52000 and 60000). However, anti-LE does not bind to proteins from leaf nuclei. Of these four tissues, leaf is the only one that does not have actively dividing cells. This observation suggests that at least some SC proteins are related to nuclear proteins from mitotically active cells.
