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Kenney, L. J.

Publications and source records attributed to Kenney, L. J..

2 recordsLinked to original sources

INTS13 Mutations Causing a Developmental Ciliopathy Disrupt Integrator Complex Assembly

Oral-facial-digital syndromes (OFD) are a heterogeneous group of congenital disorders characterized by malformations of the face and oral cavity, and digit anomalies. To date, mutations in 12 ciliary-related genes have been identified that cause several types of OFD, suggesting that OFDs constitute a subgroup of developmental ciliopathies. Through homozygosity mapping and exome sequencing of two families with variable OFD type 2, we identified distinct germline mutations in INTS13, a subunit of the Integrator complex. This 14-component complex associates with RNAPII and can cleave nascent RNA to modulate gene expression. We determined that INTS13 utilizes a discrete domain within its C-terminus to bind the Integrator cleavage module, which is disrupted by the identified germline INTS13 mutations. Depletion of INTS13 disrupts ciliogenesis in human cultured cells and causes dysregulation of a broad collection of ciliary genes. Accordingly, its knockdown in Xenopus embryos lead to motile cilia anomalies. Altogether, we show that mutations in INTS13 cause an autosomal recessive ciliopathy, which reveals key interactions within Integrator components.

biochemistry

Formin FHOD1 regulates the size of EPEC pedestals

Enteropathogenic E. coli (EPEC) is an extracellular pathogen that causes polymerization of actin filaments at the site of bacterial attachment, referred to as actin pedestals. Actin polymerization in the pedestal was believed to be solely regulated via the Nck-WASp-Arp2/3 pathway before formins were recently discovered to be associated with pedestals. Herein, we explored the collaborative role of formins in contributing to EPEC pedestal formation. In particular, we discovered that the formin FHOD1 preferentially localized to the pedestal base and its knockdown drastically reduced pedestal surface area. The pedestal localization of formin FHOD1 was found to be dependent on Tir phosphorylation at Y474, and on FHOD1 phosphorylation at Y99 from host Src family kinases (SFKs). Interestingly, differences in Arp2/3 and FHOD1 dynamics were observed. In large pedestals, Arp3 was nearly absent, but FHOD1 levels were high, suggesting that Arp2/3 and formins were segregated temporally. In line with this observation, as the pedestals grew in size, FHOD1 localization increased, while Arp3 localization decreased along the pedestals. Together, our results suggest that EPEC employs multiple actin nucleators that act at different stages of pedestal formation. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=171 SRC="FIGDIR/small/149344v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@1711602org.highwire.dtl.DTLVardef@15c060dorg.highwire.dtl.DTLVardef@914447org.highwire.dtl.DTLVardef@119cf28_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology