# Genetic Analysis of Centrioles and Cilia

> **NIH NIH R35** · WASHINGTON UNIVERSITY · 2022 · $393,750

## Abstract

Project Summary
Cilia and centrioles are evolutionarily conserved organelles that require over 2000 proteins for their
assembly and function. We use the unicellular alga, Chlamydomonas, to study these organelles.
Chlamydomonas allows us to use both haploid and diploid mutant strains obtained from unbiased forward
genetic screens together with outstanding genomics, biochemistry and microscopy to gain knowledge about
these organelles. The conservation of these organelles allows us to translate our findings to other
organisms where it is harder to do biochemistry and or genetic screens. The goal of my lab is to
understand how these organelles are built and function. As we and others have shown, ciliopathies can
exhibit a wide range of phenotypes in people that include retinal degeneration, polydactyly, cystic kidneys,
diabetes, obesity, respiratory clearance defects, shortened bones, congenital heart disease and infertility.
Defects in centrioles result in microcephaly and dwarfism. Duplication of centrioles once per cell cycle is
key; many cancers have more than two centrioles and form transient multipolar spindle structures with more
than two poles that can generate aneuploidy. I propose to address three questions using genetics,
genomics, biochemistry, and microscopy. We want to know how inner dynein arms are correctly
placed. How do they find the right address in the cilia? How are inner dynein arms assembled in
the cytoplasm? Using genes that we identified in patients with primary ciliary dyskinesia, we are
examining proteins that are needed to first assemble and then dock the large megadalton inner dynein arms
at the right address so that a functional waveform is produced. We will also perform a large-scale mating
scheme to ask if digenic inheritance can identify genes involved in dynein arm assembly and function. We
will use proteomics and cryo-EM tomography to characterize these mutations. We want to explore new
and novel pathways for regulating the precise duplication of centrioles; what redundancies are
present to guarantee only two centrioles per cell? We think that cells have layers of control of centriole
duplication. We will determine if splice site isoforms of centriolar proteins sequester other centriolar
proteins to regulate duplication. We will use proximity mapping to ask if the mother centriole provides
another level of regulation via a unique licensing site on specific triplet microtubules. We want to examine
the role of the ciliary necklace, which is a unique membrane compartment at the transition zone;
does it may play a role in membrane protein trafficking and/or in production of extracellular
vesicles? Using mutants that have a greatly reduced ciliary necklace, we will ask if extracellular vesicle
production is altered and what proteins make up the cargo of these vesicles.

## Key facts

- **NIH application ID:** 10414933
- **Project number:** 5R35GM131909-04
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** SUSAN K DUTCHER
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $393,750
- **Award type:** 5
- **Project period:** 2019-08-15 → 2024-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10414933

## Citation

> US National Institutes of Health, RePORTER application 10414933, Genetic Analysis of Centrioles and Cilia (5R35GM131909-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10414933. Licensed CC0.

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