Project Title

Identification of a novel C. elegans CAMSAP/patronin allele via RNAseq

Presenters

Academic department under which the project should be listed

CSM - Molecular and Cellular Biology

Faculty Sponsor Name

Dr. Martin Hudson

Abstract (300 words maximum)

cnd-1(ju29) mutants contain a novel ptrn-1 allele that was discovered in the cnd-1 mutant strain received from the Caenorhabditis elegans Stock Center. The ptrn-1/patronin gene is significantly down-regulated in cnd-1(ju29) mutant. We used DEXseq to map individual RNAseq reads onto a genomic scaffold. We find that cnd- 1(ju29) mutants have significant differences in transcript levels at the 3’ end and 3’ untranslated region of the ptrn-1/patronin gene. ptrn-1 codes for a Ca2+ sensitive microtubule minus-end binding protein and is an ortholog of human CAMSAP (Patronin/Nezha/Calmodulin- and spectrin-associated protein). The CKK (calmodulin-regulated spectrin associated domain) is located at the C-terminus in CAMSAP proteins. ptrn-1 inhibits DLK-1 and the MAPK cascade and it is suggested that it inhibits microtubule dynamics. The CKK domain of ptrn-1 is necessary and sufficient for its function in MT dynamics. This allele is a 7kb deletion that removes the 3’UTR of the ptrn-1 gene, that appears to affect transcript levels of the 3’ coding region also. Does this novel ptrn-1 allele enhance cnd-1(ju29) axon guidance defects? Why a ptrn-1 gene that is not on the same chromosome as cnd-1 should co-segregate is not known. We hypothesize that ptrn-1 mutants enhance the axon guidance defects seen in cnd-1 mutants. Work is on-going to cross our novel ptrn-1 allele into a different cnd-1 allele (gk718) to assay for axon guidance effects. Then a PCR of F2 generation will be performed to confirm outcross of ptrn- 1(-/-) away from cnd-1(ju29). The DNA will then be sent for analysis.

Project Type

Oral Presentation (15-min time slots)

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Identification of a novel C. elegans CAMSAP/patronin allele via RNAseq

cnd-1(ju29) mutants contain a novel ptrn-1 allele that was discovered in the cnd-1 mutant strain received from the Caenorhabditis elegans Stock Center. The ptrn-1/patronin gene is significantly down-regulated in cnd-1(ju29) mutant. We used DEXseq to map individual RNAseq reads onto a genomic scaffold. We find that cnd- 1(ju29) mutants have significant differences in transcript levels at the 3’ end and 3’ untranslated region of the ptrn-1/patronin gene. ptrn-1 codes for a Ca2+ sensitive microtubule minus-end binding protein and is an ortholog of human CAMSAP (Patronin/Nezha/Calmodulin- and spectrin-associated protein). The CKK (calmodulin-regulated spectrin associated domain) is located at the C-terminus in CAMSAP proteins. ptrn-1 inhibits DLK-1 and the MAPK cascade and it is suggested that it inhibits microtubule dynamics. The CKK domain of ptrn-1 is necessary and sufficient for its function in MT dynamics. This allele is a 7kb deletion that removes the 3’UTR of the ptrn-1 gene, that appears to affect transcript levels of the 3’ coding region also. Does this novel ptrn-1 allele enhance cnd-1(ju29) axon guidance defects? Why a ptrn-1 gene that is not on the same chromosome as cnd-1 should co-segregate is not known. We hypothesize that ptrn-1 mutants enhance the axon guidance defects seen in cnd-1 mutants. Work is on-going to cross our novel ptrn-1 allele into a different cnd-1 allele (gk718) to assay for axon guidance effects. Then a PCR of F2 generation will be performed to confirm outcross of ptrn- 1(-/-) away from cnd-1(ju29). The DNA will then be sent for analysis.