PUBLICATION

Resting cells rely on the DNA helicase component MCM2 to build cilia

Authors

Casar Tena, T., Maerz, L.D., Szafranski, K., Groth, M., Blätte, T.J., Donow, C., Matysik, S., Walther, P., Jeggo, P.A., Burkhalter, M.D., Philipp, M.

ID

ZDB-PUB-181018-8

Date

2018

Source

Nucleic acids research 47(1): 134-151 (Journal)

Registered Authors

Burkhalter, Martin, Philipp, Melanie

Keywords

none

MeSH Terms

Animals
Mitosis/genetics
Humans
Zebrafish/genetics
Ciliopathies/genetics
Ciliopathies/pathology
Cilia/genetics*
Cilia/pathology
Transcription Initiation Site
DNA Helicases/genetics*
Transcription, Genetic*
Minichromosome Maintenance Complex Component 2/genetics*
Minichromosome Maintenance Complex Component 7/genetics*

(all 13)

PubMed

30329080 Full text @ Nucleic Acids Res.

Abstract

Minichromosome maintenance (MCM) proteins facilitate replication by licensing origins and unwinding the DNA double strand. Interestingly, the number of MCM hexamers greatly exceeds the number of firing origins suggesting additional roles of MCMs. Here we show a hitherto unanticipated function of MCM2 in cilia formation in human cells and zebrafish that is uncoupled from replication. Zebrafish depleted of MCM2 develop ciliopathy-phenotypes including microcephaly and aberrant heart looping due to malformed cilia. In non-cycling human fibroblasts, loss of MCM2 promotes transcription of a subset of genes, which cause cilia shortening and centriole overduplication. Chromatin immunoprecipitation experiments show that MCM2 binds to transcription start sites of cilia inhibiting genes. We propose that such binding may block RNA polymerase II-mediated transcription. Depletion of a second MCM (MCM7), which functions in complex with MCM2 during its canonical functions, reveals an overlapping cilia-deficiency phenotype likely unconnected to replication, although MCM7 appears to regulate a distinct subset of genes and pathways. Our data suggests that MCM2 and 7 exert a role in ciliogenesis in post-mitotic tissues.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
gli1	WT	standard conditions	Prim-5	whole organism	RTPCR	Fig. 3
gli1	WT + MO4-mcm2	standard conditions	Prim-5	whole organism	RTPCR	Fig. 3
ins	WT	standard conditions	Long-pec	endocrine pancreas	ISH	Fig. 2
ins	WT + MO3-mcm2	standard conditions	Long-pec	endocrine pancreas	ISH	Fig. 2
ins	WT + MO3-mcm2	standard conditions	Long-pec	whole organism	ISH	Fig. 2
ins	WT + MO4-mcm2	standard conditions	Long-pec	endocrine pancreas	ISH	Fig. 2
mcm2	WT	standard conditions	5-9 somites	hindbrain neural rod	ISH	Fig. 3
mcm2	WT	standard conditions	5-9 somites	midbrain neural rod	ISH	Fig. 3
mcm2	WT	standard conditions	5-9 somites	optic vesicle	ISH	Fig. 3
mcm2	WT	standard conditions	1-4 somites	paraxial mesoderm	ISH	Fig. 3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment by environment: 1-[4-fluoro-3-(trifluoromethyl)phenyl]-3-(5-pyridin-4-yl-1,3,4-thiadiazol-2-yl)urea	20-25 somites	lateral plate mesoderm determination of left/right asymmetry in lateral mesoderm occurrence, normal	Fig. 2
WT + MO1-mcm7	standard conditions	5-9 somites	Kupffer's vesicle area, normal	Fig. 9
WT + MO1-mcm7	standard conditions	5-9 somites	Kupffer's vesicle motile cilium decreased length, abnormal	Fig. 9
WT + MO1-mcm7	standard conditions	20-25 somites	lateral plate mesoderm determination of left/right asymmetry in lateral mesoderm decreased occurrence, abnormal	Fig. 9
WT + MO1-mcm7	standard conditions	Prim-5	whole organism mcm7 expression decreased amount, abnormal	Fig. 8
WT + MO1-mcm7	standard conditions	Long-pec	eye decreased size, abnormal	Fig. 8
WT + MO1-mcm7	standard conditions	Long-pec	head decreased size, abnormal	Fig. 8
WT + MO1-mcm7	standard conditions	Long-pec	heart edematous, abnormal	Fig. 8
WT + MO3-mcm2	standard conditions	5-9 somites	Kupffer's vesicle area, normal	Fig. 3
WT + MO3-mcm2	standard conditions	5-9 somites	Kupffer's vesicle has normal numbers of parts of type Kupffer's vesicle motile cilium, normal	Fig. 3

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Mutations / Transgenics

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
mcm2	MO3-mcm2	MRPHLNO
mcm2	MO4-mcm2	MRPHLNO
mcm7	MO1-mcm7	MRPHLNO

Fish

Fish
WT
WT + MO1-mcm7
WT + MO3-mcm2
WT + MO4-mcm2

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-cetn1	polyclonal		IgG	Rabbit
Ab1-prkcz	polyclonal	prkci prkcz	IgG	Rabbit
Ab1-tuba	monoclonal		IgG2b	Mouse
Ab3-pcnt	polyclonal			Rabbit

Orthology

Engineered Foreign Genes

Mapping

Casar Tena et al., 2018 ZDB-PUB-181018-8 PMID:30329080

Resting cells rely on the DNA helicase component MCM2 to build cilia

Casar Tena et al., 2018

ZDB-PUB-181018-8
PMID:30329080