| 2000 |
NKLAM (RNF19B) encodes a zinc finger protein that resides in NK cytolytic granules; antisense oligonucleotide-mediated knockdown of NKLAM inhibits cytotoxic function of NK cells and CTLs, establishing a direct role in cellular cytotoxicity. Two human isoforms (587 and 731 residue proteins) arise from alternative splicing, with the larger form predominant. |
Antisense oligonucleotide knockdown of NKLAM in NK cells/CTLs with cytotoxicity assays; cDNA cloning, genomic structure analysis, and expression profiling |
Immunogenetics |
Medium |
10912506
|
| 2012 |
NKLAM is a phagosomal E3 ubiquitin ligase in macrophages; it translocates to the phagosome early during maturation coinciding with elevated phagosomal ubiquitinated proteins, co-localizes with ingested E. coli, and NKLAM-deficient macrophages show reduced bacterial killing. |
Confocal microscopy (co-localization with E. coli and IgG-opsonized latex beads), subcellular fractionation, bacterial killing assays with NKLAM-KO bone marrow-derived macrophages vs. wild type |
Cellular immunology |
Medium |
23085241
|
| 2014 |
NKLAM positively regulates iNOS expression and NO production in LPS-stimulated macrophages by promoting STAT1 tyrosine phosphorylation, IFNβ production, and NFκB p65 nuclear translocation; NKLAM-KO macrophages show reduced p65 expression, defective p65 phosphorylation at S536, and diminished NFκB transcriptional activity independent of IKBα degradation. |
NKLAM-KO vs. WT bone marrow-derived macrophage comparisons: ELISA (NO, IFNβ), Western blot (iNOS, phospho-STAT1, p65, phospho-p65, IKBα), confocal microscopy (p65 nuclear translocation), NFκB luciferase reporter transfection assay |
Immunobiology |
Medium |
25182373
|
| 2016 |
NKLAM mediates K63-linked ubiquitination of STAT1 in macrophages; NKLAM transiently localizes to the IFNγ receptor complex during IFNγ stimulation, binds STAT1, and this ubiquitination is required for STAT1 DNA-binding to gamma activation sequences and full STAT1-mediated transcription. Loss of NKLAM results in hyperphosphorylation of JAK1 and STAT1 but impaired STAT1–DNA binding, without affecting STAT1 nuclear translocation. |
Co-immunoprecipitation (NKLAM with IFNγ receptor complex and STAT1), ubiquitination assay (K63-linkage specificity), confocal microscopy (NKLAM localization), EMSA (STAT1-DNA binding), NKLAM-KO macrophage nucleofection with STAT1-driven luciferase reporter, RT-PCR (STAT1 target gene mRNA) |
Cellular signalling |
High |
27570112
|
| 2018 |
In vivo, NKLAM-KO mice infected with S. pneumoniae show higher bacterial lung burden, reduced STAT1 and STAT3 phosphorylation, and impaired neutrophil and NK cell recruitment; NKLAM-KO neutrophils and macrophages are individually defective in killing S. pneumoniae, confirming NKLAM's positive regulatory role in bactericidal activity in multiple innate immune cell types. |
Inhalation infection model with NKLAM-KO and WT mice; bacterial CFU enumeration, flow cytometry (lung immune cell populations), Western blot (phospho-STAT1, phospho-STAT3), ELISA (cytokines), ex vivo killing assays with isolated neutrophils and macrophages |
PloS one |
Medium |
29518136
|
| 2019 |
NKLAM regulates antiviral innate immunity during Sendai virus infection: NKLAM-KO mice show reduced STAT1 and NFκB p65 phosphorylation, attenuated autophagy (reduced LC3 and p62/SQSTM1), fewer lung neutrophils and macrophages, diminished pro-inflammatory cytokine production, and paradoxically attenuated viral replication at low dose but increased susceptibility at high dose. |
In vivo Sendai virus infection model with NKLAM-KO vs. WT mice; weight monitoring, viral titer, flow cytometry, Western blot (phospho-STAT1, phospho-NFκB p65, LC3, p62), ELISA (cytokines) |
PloS one |
Medium |
31539400
|
| 2023 |
RNF19B acts as the E3 ubiquitin ligase that mediates polyubiquitination and proteasomal degradation of RAC1 in NSCLC cells; DIRAS3 promotes the physical interaction between RNF19B and RAC1, enhancing RAC1 degradation and suppressing cell migration via the DIRAS3–RNF19B–RAC1 axis. |
Co-immunoprecipitation (RNF19B–RAC1 interaction with and without DIRAS3), ubiquitination assay (polyubiquitination of RAC1), proteasome inhibitor rescue of RAC1 levels, cell migration assays, knockdown/overexpression of DIRAS3 and RNF19B |
iScience |
Medium |
37485351
|
| 2025 |
Cysteine 301 of NKLAM is the catalytic residue essential for its RBR E3 ubiquitin ligase activity; C301A and C301S mutations abrogate ligase activity. Wild-type but not C301A NKLAM inhibits cellular proliferation and facilitates proteasomal degradation of c-Myc (reducing c-Myc half-life from 27 to 12 min). Prolonged NKLAM expression induces apoptosis; the C301S mutant induces apoptosis comparably to wild-type despite lacking ligase activity, indicating an alternative, ligase-independent apoptotic pathway. |
Site-directed mutagenesis (C301A, C301S), inducible expression in HEK293 and K562 cells, cell proliferation assays, metabolic activity assays, cycloheximide chase (c-Myc half-life), proteasome inhibitor rescue, annexin-V staining, caspase activation assays |
The Journal of biological chemistry |
High |
40273985
|
| 2025 |
RNF19B, whose expression is transcriptionally promoted by the zinc-finger transcription factor ZNF573, mediates ubiquitination of PIK3CA in prostate cancer cells; loss of ZNF573 (via promoter hypermethylation) reduces RNF19B expression and impairs PIK3CA ubiquitination, enhancing PCa proliferation and invasion. |
ZNF573 overexpression/knockdown in PCa cells in vitro and in vivo, chromatin immunoprecipitation or luciferase reporter (ZNF573 transactivation of RNF19B), co-immunoprecipitation and ubiquitination assay (RNF19B–PIK3CA), proliferation and invasion assays |
Oncogene |
Medium |
40973794
|