Nuclear FAM21 participates in NF-κB-dependent gene regulation in pancreatic cancer cells

Zhi Hui Deng, Timothy S. Gomez, Douglas G. Osborne, Christine A. Phillips-Krawczak, Jin San Zhang, Daniel D Billadeau

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The pentameric WASH complex is best known for its role in regulating receptor trafficking from retromer-rich endosomal subdomains. FAM21 functions to stabilize the WASH complex through its N-terminal head domain and localizes it to endosomes by directly binding the retromer through its extended C-terminal tail. Herein, we used affinity purification combined with mass spectrometry to identify additional FAM21-interacting proteins. Surprisingly, multiple components of the nuclear factor κB (NF-κB) pathway were identified, including the p50 and p65 (RelA) NF-κB subunits. We show that FAM21 interacts with these components and regulates NF-κB-dependent gene transcription at the level of p65 chromatin binding. We further demonstrate that FAM21 contains a functional monopartite nuclear localization signal sequence (NLS) as well as a CRM1/exportin1-dependent nuclear export signal (NES), both of which work jointly with the N-terminal head domain and C-terminal retromer recruitment domain to regulate FAM21 cytosolic and nuclear subcellular localization. Finally, our findings indicate that FAM21 depletion sensitizes pancreatic cancer cells to gemcitabine and 5-fluorouracil. Thus, FAM21 not only functions as an integral component of the cytoplasmic WASH complex, but also modulates NF-κB gene transcription in the nucleus.

Original languageEnglish (US)
Pages (from-to)373-384
Number of pages12
JournalJournal of Cell Science
Volume128
Issue number2
DOIs
StatePublished - 2015

Fingerprint

gemcitabine
Pancreatic Neoplasms
Head
Nuclear Export Signals
Nuclear Localization Signals
Endosomes
Protein Sorting Signals
Fluorouracil
Genes
Chromatin
Mass Spectrometry
Proteins

Keywords

  • FAM21
  • NF-κB
  • Pancreatic cancer
  • Retromer
  • WASH complex

ASJC Scopus subject areas

  • Cell Biology

Cite this

Deng, Z. H., Gomez, T. S., Osborne, D. G., Phillips-Krawczak, C. A., Zhang, J. S., & Billadeau, D. D. (2015). Nuclear FAM21 participates in NF-κB-dependent gene regulation in pancreatic cancer cells. Journal of Cell Science, 128(2), 373-384. https://doi.org/10.1242/jcs.161513

Nuclear FAM21 participates in NF-κB-dependent gene regulation in pancreatic cancer cells. / Deng, Zhi Hui; Gomez, Timothy S.; Osborne, Douglas G.; Phillips-Krawczak, Christine A.; Zhang, Jin San; Billadeau, Daniel D.

In: Journal of Cell Science, Vol. 128, No. 2, 2015, p. 373-384.

Research output: Contribution to journalArticle

Deng, ZH, Gomez, TS, Osborne, DG, Phillips-Krawczak, CA, Zhang, JS & Billadeau, DD 2015, 'Nuclear FAM21 participates in NF-κB-dependent gene regulation in pancreatic cancer cells', Journal of Cell Science, vol. 128, no. 2, pp. 373-384. https://doi.org/10.1242/jcs.161513
Deng, Zhi Hui ; Gomez, Timothy S. ; Osborne, Douglas G. ; Phillips-Krawczak, Christine A. ; Zhang, Jin San ; Billadeau, Daniel D. / Nuclear FAM21 participates in NF-κB-dependent gene regulation in pancreatic cancer cells. In: Journal of Cell Science. 2015 ; Vol. 128, No. 2. pp. 373-384.
@article{86c6224b2b8e41f896dde44a77829198,
title = "Nuclear FAM21 participates in NF-κB-dependent gene regulation in pancreatic cancer cells",
abstract = "The pentameric WASH complex is best known for its role in regulating receptor trafficking from retromer-rich endosomal subdomains. FAM21 functions to stabilize the WASH complex through its N-terminal head domain and localizes it to endosomes by directly binding the retromer through its extended C-terminal tail. Herein, we used affinity purification combined with mass spectrometry to identify additional FAM21-interacting proteins. Surprisingly, multiple components of the nuclear factor κB (NF-κB) pathway were identified, including the p50 and p65 (RelA) NF-κB subunits. We show that FAM21 interacts with these components and regulates NF-κB-dependent gene transcription at the level of p65 chromatin binding. We further demonstrate that FAM21 contains a functional monopartite nuclear localization signal sequence (NLS) as well as a CRM1/exportin1-dependent nuclear export signal (NES), both of which work jointly with the N-terminal head domain and C-terminal retromer recruitment domain to regulate FAM21 cytosolic and nuclear subcellular localization. Finally, our findings indicate that FAM21 depletion sensitizes pancreatic cancer cells to gemcitabine and 5-fluorouracil. Thus, FAM21 not only functions as an integral component of the cytoplasmic WASH complex, but also modulates NF-κB gene transcription in the nucleus.",
keywords = "FAM21, NF-κB, Pancreatic cancer, Retromer, WASH complex",
author = "Deng, {Zhi Hui} and Gomez, {Timothy S.} and Osborne, {Douglas G.} and Phillips-Krawczak, {Christine A.} and Zhang, {Jin San} and Billadeau, {Daniel D}",
year = "2015",
doi = "10.1242/jcs.161513",
language = "English (US)",
volume = "128",
pages = "373--384",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "2",

}

TY - JOUR

T1 - Nuclear FAM21 participates in NF-κB-dependent gene regulation in pancreatic cancer cells

AU - Deng, Zhi Hui

AU - Gomez, Timothy S.

AU - Osborne, Douglas G.

AU - Phillips-Krawczak, Christine A.

AU - Zhang, Jin San

AU - Billadeau, Daniel D

PY - 2015

Y1 - 2015

N2 - The pentameric WASH complex is best known for its role in regulating receptor trafficking from retromer-rich endosomal subdomains. FAM21 functions to stabilize the WASH complex through its N-terminal head domain and localizes it to endosomes by directly binding the retromer through its extended C-terminal tail. Herein, we used affinity purification combined with mass spectrometry to identify additional FAM21-interacting proteins. Surprisingly, multiple components of the nuclear factor κB (NF-κB) pathway were identified, including the p50 and p65 (RelA) NF-κB subunits. We show that FAM21 interacts with these components and regulates NF-κB-dependent gene transcription at the level of p65 chromatin binding. We further demonstrate that FAM21 contains a functional monopartite nuclear localization signal sequence (NLS) as well as a CRM1/exportin1-dependent nuclear export signal (NES), both of which work jointly with the N-terminal head domain and C-terminal retromer recruitment domain to regulate FAM21 cytosolic and nuclear subcellular localization. Finally, our findings indicate that FAM21 depletion sensitizes pancreatic cancer cells to gemcitabine and 5-fluorouracil. Thus, FAM21 not only functions as an integral component of the cytoplasmic WASH complex, but also modulates NF-κB gene transcription in the nucleus.

AB - The pentameric WASH complex is best known for its role in regulating receptor trafficking from retromer-rich endosomal subdomains. FAM21 functions to stabilize the WASH complex through its N-terminal head domain and localizes it to endosomes by directly binding the retromer through its extended C-terminal tail. Herein, we used affinity purification combined with mass spectrometry to identify additional FAM21-interacting proteins. Surprisingly, multiple components of the nuclear factor κB (NF-κB) pathway were identified, including the p50 and p65 (RelA) NF-κB subunits. We show that FAM21 interacts with these components and regulates NF-κB-dependent gene transcription at the level of p65 chromatin binding. We further demonstrate that FAM21 contains a functional monopartite nuclear localization signal sequence (NLS) as well as a CRM1/exportin1-dependent nuclear export signal (NES), both of which work jointly with the N-terminal head domain and C-terminal retromer recruitment domain to regulate FAM21 cytosolic and nuclear subcellular localization. Finally, our findings indicate that FAM21 depletion sensitizes pancreatic cancer cells to gemcitabine and 5-fluorouracil. Thus, FAM21 not only functions as an integral component of the cytoplasmic WASH complex, but also modulates NF-κB gene transcription in the nucleus.

KW - FAM21

KW - NF-κB

KW - Pancreatic cancer

KW - Retromer

KW - WASH complex

UR - http://www.scopus.com/inward/record.url?scp=84921325996&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84921325996&partnerID=8YFLogxK

U2 - 10.1242/jcs.161513

DO - 10.1242/jcs.161513

M3 - Article

C2 - 25431135

AN - SCOPUS:84921325996

VL - 128

SP - 373

EP - 384

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 2

ER -