Phosphorylation of CEACAM1 Molecule by Calmodulin Kinase IID in a Three-dimensional Model of Mammary Gland Lumen Formation

Chen, Charng-Jui; Shively, John E.

doi:10.1074/jbc.m113.496992

Cited by 17 publications

(16 citation statements)

References 30 publications

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“…Thus, elevation of intracellular calcium as a consequence of activation via a variety of cell-specific receptors provides the impetus for calmodulin binding and CEACAM1 monomerization [71]. This consensus sequence also is required for lumen formation in transformed breast epithelial cells through calmodulin kinase IID (CaMKIID)-mediated phosphorylation of the shared threonine (T448) [72,73]. Finally, this consensus motif accounts for the ability of both CEACAM1-L and CEACAM1-S to bind subdomain 3 of actin which is mediated by residues F445 and K447 [72,73].…”

Section: The Cytoplasmic Domain Determines Inhibitory or Non-inhibitomentioning

confidence: 99%

CEACAM1 structure and function in immunity and its therapeutic implications

Kim

Huang

Gandhi

et al. 2019

Seminars in Immunology

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The type I membrane protein receptor carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) distinctively exhibits significant alternative splicing that allows for tunable functions upon homophilic binding. CEACAM1 is highly expressed in the tumor environment and is strictly regulated on lymphocytes such that its expression is restricted to activated cells where it is now recognized to function in tolerance pathways. CEACAM1 is also an important target for microbes which have co-opted these attributes of CEACAM1 for the purposes of invading the host and evading the immune system. These properties, among others, have focused attention on CEACAM1 as a unique target for immunotherapy in autoimmunity and cancer. This review examines recent structural information derived from the characterization of CEACAM1:CEACAM1 interactions and heterophilic modes of binding especially to microbes and how this relates to CEACAM1 function. Through this, we aim to provide insights into targeting CEACAM1 for therapeutic intervention.

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Section: The Cytoplasmic Domain Determines Inhibitory or Non-inhibitomentioning

confidence: 99%

CEACAM1 structure and function in immunity and its therapeutic implications

Kim

Huang

Gandhi

et al. 2019

Seminars in Immunology

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“…injection of LPS for 2 hours, Liver, spleen and duodenum of both WT and Ceacam1 -/mice were harvested separately, homogenized and lysed in 1% NP-40 lysis buffer as previously described 64 VAV1 pAb, SHP1 pAb, phospho-SHP-1 Y564 pAb, Src, and 4G10 pAb from Cell Signaling Technology, Inc., Danvers, MA; anti-β-actin mAb from GeneTex, Inc., Irvine, CA) and infraredlabeled IRDye secondary antibodies. Detection was carried out using the Odyssey infrared imaging 77 .…”

Section: Immunoblot Analysis and Immunoprecipitation (Ip)mentioning

confidence: 99%

CEACAM1 regulates the IL-6 mediated fever response to LPS through the RP105 receptor in murine monocytes

Zhang

Placa

Kujawski

et al. 2018

Preprint

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short title: CEACAM1 regulation of monocyte IL6 response to LPS ABSTRACT Systemic inflammation and the fever response to pathogens are coordinately regulated by IL-6 and IL-1β. We previously showed that CEACAM1 regulates the LPS driven expression of IL-1β in murine neutrophils through its ITIM receptor. We now show that the prompt secretion of IL-6 in response to LPS is regulated by CEACAM1 expression on bone marrow monocytes.Ceacam1 -/mice over-produce IL-6 in response to an i.p. LPS challenge, resulting in prolonged surface temperature depression and overt diarrhea compared to their wild type counterparts.Intraperitoneal injection of a 64 Cu-labeled LPS, PET imaging agent shows confined localization to the peritoneal cavity, and fluorescent labeled LPS is taken up by myeloid splenocytes and muscle endothelial cells. While bone marrow monocytes and their progenitors (CD11b + Ly6G -) express IL-6 in the early response (<2 hours) to LPS in vitro, these cells are not detected in the bone marrow after in vivo LPS treatment due to their rapid and complete mobilization to the periphery. Notably, tissue macrophages are not involved in the early IL-6 response to LPS. In contrast to human monocytes, TLR4 is not expressed on murine bone marrow monocytes.Instead, the alternative LPS receptor RP105 is expressed and recruits MD1, CD14, Src, VAV1 and β-actin in response to LPS to produce IL-6. CEACAM1 negatively regulates RP105 signaling in monocytes by recruitment of SHP-1, resulting in the sequestration of pVAV1 and βactin from RP105. This novel pathway and regulation of IL-6 producing by CEACAM1 defines a novel role for monocytes in the fever of mice to LPS. (250 words) AUTHOR SUMMARYFever is one of the most common signs of the immune response to pathogens. The fever response to LPS or endotoxin of gram-negative bacteria is mediated by the combined action of two cytokines, IL-1β and IL-6. Regulation of their production in response to LPS is an important area of investigation. While we previously showed that the regulation of IL-1β production in neutrophils is through the lymphocyte receptor CEACAM1, we were interested if a similar mechanism operated for IL-6. Using a mouse model in which the CEACAM1 gene was knocked out, we show that IL-6 is over-produced compared to normal mice, and that monocytes, rather than neutrophils were the principal IL-6 producing cells. Surprisingly, murine monocytes do not express TLR4, the most commonly studied receptor for LPS, but instead express the low affinity LPS receptor, RP105, a receptor common expressed on B-cells.Furthermore, we show that bone marrow monocytes are rapidly released into the blood and home to tissues throughout the body in response to LPS. These findings explain much of the confusion in the literature concerning the immediate source of IL-6 and the distinct differences between murine and human monocytes in their in responses to LPS. (word count 196)

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“…Due to its small size it was possible to mutate each of the 12 amino acids and determine which were critical for lumen formation. We identified critical amino acids Phe-454 and Thr-457, the former for interaction with actin and calmodulin (6,7), and the latter for phosphorylation by calmodulin kinase 2D (8). Although these studies indicated that the cytoplasmic domain of CEACAM1-SF interacted with the cytoskeleton, they did not reveal which downstream genes were, in turn, regulated to enable lumen formation.…”

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confidence: 72%

miRNA-342 Regulates CEACAM1-induced Lumen Formation in a Three-dimensional Model of Mammary Gland Morphogenesis

Weng

Shively

2016

Journal of Biological Chemistry

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Lumen formation of breast epithelium is rapidly lost during tumorigenesis along with expression of cell adhesion molecule CEACAM1. CEACAM1 induces lumena in a three-dimensional culture of MCF7/CEACAM1 cells that otherwise fail to form lumena. We hypothesized miRNAs may be involved because >400 genes were up-or down-regulated in MCF7/CEACAM1 cells and miRNAs may modify global expression patterns. Comparative analysis of miRNA expression in MCF7 versus MCF7/ CEACAM1 cells revealed two miRNAs significantly down-regulated (hsa-miR-30a-3p by 6.73-fold and hsa-miR-342-5p by 5.68-fold). Location of miR-342 within an intron of the EVL gene, hypermethylated and involved in tumorigenesis, suggested that miR-342 overexpression may block lumen formation. In fact, overexpression of miR-342 in MCF7/CEACAM1 cells significantly blocked lumen formation (p < 0.001). ID4, a dominant-negative inhibitor of basic helix-loop-helix transcription factors, up-regulated in MCF7/CEACAM1 cells, down-regulated in breast cancer, and containing a miR-342 binding site, was tested as a potential target of miR-342. The ratio of ID4 to miR-342 increased from 1:2 in MCF7 cells to 30:1 in MCF7/ CEACAM1 cells and a miR-342 inhibitor was able to induce 3-UTR ID4 reporter activity in MCF7 cells. Because 5-methylcytosine methyltransferase DNMT1 is also a potential target of miR-342, we inhibited miR-342 in MCF7 cells and found DNMT1 was up-regulated with no change in EVL expression, suggesting that miR-342 regulates DNMT1 expression but DNMT1 does not affect the EVL expression in these cells. We conclude that the regulation of lumen formation by miR-342 involves at least two of its known targets, namely ID4 and DNMT1.Lumen formation of mammary epithelial cells in a threedimensional culture is an in vitro assay that phenocopies mammary epithelial gland formation (1-3). The loss of lumen formation as exhibited by most, if not all, breast cancer cells in the three-dimensional culture assay phenocopies the premalignant stage of breast cancer know as ductal carcinoma in situ. One approach to identify genes that may govern the change from normal to (pre)malignancy is to transfect breast cancer cells with genes down-regulated in breast cancer and assay their regain of function in the lumen formation assay. A second approach is to block suspect gene expression in breast cancer cells for regain of function, or block gene expression in normal breast epithelial cells for loss of function. One gene that stands out in these assays is the cell-cell adhesion molecule CEACAM1 that is highly expressed in normal breast with a luminal expression pattern (4, 5), and is consistently down-regulated in premalignant and invasive breast cancer (4, 5).Previously, we have shown that transfection of CEACAM1 into MCF7 breast cancer cells resulted in lumen formation when the cells were cultured three-dimensionally (6), whereas anti-CEACAM1 antibodies blocked lumen formation of the normal breast epithelial cell line MCF10F that express CEACAM1 (5). Although CEACAM1 is expressed as mu...

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Phosphorylation of CEACAM1 Molecule by Calmodulin Kinase IID in a Three-dimensional Model of Mammary Gland Lumen Formation

Cited by 17 publications

References 30 publications

CEACAM1 structure and function in immunity and its therapeutic implications

CEACAM1 structure and function in immunity and its therapeutic implications

CEACAM1 regulates the IL-6 mediated fever response to LPS through the RP105 receptor in murine monocytes

miRNA-342 Regulates CEACAM1-induced Lumen Formation in a Three-dimensional Model of Mammary Gland Morphogenesis

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