TY - JOUR
T1 - Sterile α-motif domain requirement for cellular signaling and survival
AU - Ray, Suhita
AU - Chee, Linda
AU - Matson, Daniel R.
AU - Palermo, Nick Y.
AU - Bresnick, Emery H.
AU - Hewitt, Kyle J.
N1 - Funding Information:
This work was supported by National Institutes of Health Grants K01DK113117 (to K. J. H.), Nebraska Stem Cell Research Grant DHHS 2019-01, DK50107 (to E. H. B.), Cancer Center Support Grant P30CA014520, and the COBRE Nebraska Center for Molecular Target Discovery and Development Grant P20GM121316. The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
This work was supported by National Institutes of Health Grants K01DK113117 (to K. J. H.), Nebraska Stem Cell Research Grant DHHS 2019-01, DK50107 (to E. H. B.), Cancer Center Support Grant P30CA014520, and the COBRE Nebraska Center for Molecular Target Discovery and Develop-ment Grant P20GM121316. The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the respon-sibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2020 Ray et al. Published by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2020/5/15
Y1 - 2020/5/15
N2 - Hundreds of sterile α-motif (SAM) domains have predicted structural similarities and are reported to bind proteins, lipids, or RNAs. However, the majority of these domains have not been analyzed functionally. Previously, we demonstrated that a SAM domain-containing protein, SAMD14, promotes SCF/protooncogene c-Kit (c-Kit) signaling, erythroid progenitor function, and erythrocyte regeneration. Deletion of a Samd14 enhancer (Samd14–Enh), occupied by GATA2 and SCL/TAL1 transcription factors, reduces SAMD14 expression in bone marrow and spleen and is lethal in a hemolytic anemia mouse model. To rigorously establish whether Samd14–Enh deletion reduces anemia-dependent c-Kit signaling by lowering SAMD14 levels, we developed a genetic rescue assay in murine Samd14–Enh-/- primary erythroid precursor cells. SAMD14 expression at endogenous levels rescued c-Kit signaling. The conserved SAM domain was required for SAMD14 to increase colony-forming activity, c-Kit signaling, and progenitor survival. To elucidate the molecular determinants of SAM domain function in SAMD14, we substituted its SAM domain with distinct SAM domains predicted to be structurally similar. The chimeras were less effective than SAMD14 itself in rescuing signaling, survival, and colony-forming activities. Thus, the SAMD14 SAM domain has attributes that are distinct from other SAM domains and underlie SAMD14 function as a regulator of cellular signaling and erythrocyte regeneration.
AB - Hundreds of sterile α-motif (SAM) domains have predicted structural similarities and are reported to bind proteins, lipids, or RNAs. However, the majority of these domains have not been analyzed functionally. Previously, we demonstrated that a SAM domain-containing protein, SAMD14, promotes SCF/protooncogene c-Kit (c-Kit) signaling, erythroid progenitor function, and erythrocyte regeneration. Deletion of a Samd14 enhancer (Samd14–Enh), occupied by GATA2 and SCL/TAL1 transcription factors, reduces SAMD14 expression in bone marrow and spleen and is lethal in a hemolytic anemia mouse model. To rigorously establish whether Samd14–Enh deletion reduces anemia-dependent c-Kit signaling by lowering SAMD14 levels, we developed a genetic rescue assay in murine Samd14–Enh-/- primary erythroid precursor cells. SAMD14 expression at endogenous levels rescued c-Kit signaling. The conserved SAM domain was required for SAMD14 to increase colony-forming activity, c-Kit signaling, and progenitor survival. To elucidate the molecular determinants of SAM domain function in SAMD14, we substituted its SAM domain with distinct SAM domains predicted to be structurally similar. The chimeras were less effective than SAMD14 itself in rescuing signaling, survival, and colony-forming activities. Thus, the SAMD14 SAM domain has attributes that are distinct from other SAM domains and underlie SAMD14 function as a regulator of cellular signaling and erythrocyte regeneration.
UR - http://www.scopus.com/inward/record.url?scp=85084820781&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85084820781&partnerID=8YFLogxK
U2 - 10.1074/jbc.RA119.011895
DO - 10.1074/jbc.RA119.011895
M3 - Article
C2 - 32241909
AN - SCOPUS:85084820781
VL - 295
SP - 7113
EP - 7125
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 20
ER -