Leucine zipper transcription factor-like 1 binds adaptor protein complex-1 and 2 and participates in trafficking of transferrin receptor 1

Abstract

LZTFL1 participates in immune synapse formation, ciliogenesis, and the localization of ciliary proteins, and knockout of LZTFL1 induces abnormal distribution of heterotetrameric adaptor protein complex-1 (AP-1) in the Lztfl1-knockout mouse photoreceptor cells, suggesting that LZTFL1 is involved in intracellular transport. Here, we demonstrate that in vitro LZTFL1 directly binds to AP-1 and AP-2 and coimmunoprecipitates AP-1 and AP-2 from cell lysates. DxxFxxLxxxR motif of LZTFL1 is essential for these bindings, suggesting LZTFL1 has roles in AP-1 and AP-2-mediated protein trafficking. Since AP-1 and AP-2 are known to be involved in transferrin receptor 1 (TfR1) trafficking, the effect of LZTFL1 on TfR1 recycling was analyzed. TfR1, AP-1 and LZTFL1 from cell lysates could be coimmunoprecipitated. However, pull-down results indicate there is no direct interaction between TfR1 and LZTFL1, suggesting that LZTFL1 interaction with TfR1 is indirect through AP-1. We report the colocalization of LZTFL1 and AP-1, AP-1 and TfR1 as well as LZTFL1 and TfR1 in the perinuclear region (PNR) and the cytoplasm, suggesting a potential complex between LZTFL1, AP-1 and TfR1. The results from the disruption of adaptin recruitment with brefeldin A treatment suggested ADP-ribosylation factor-dependent localization of LZFL1 and AP-1 in the PNR. Knockdown of AP-1 reduces the level of LZTFL1 in the PNR, suggesting that AP-1 plays a role in LZTFL1 trafficking. Knockout of LZTFL1 reduces the cell surface level and the rate of internalization of TfR1, leading to a decrease of transferrin uptake, efflux, and internalization. However, knockout of LZTFL1 did not affect the cell surface levels of epidermal growth factor receptor and cation-independent mannose 6-phosphate receptor, indicating that LZTFL1 specifically regulates the cell surface level of TfR1. These data support a novel role of LZTFL1 in regulating the cell surface TfR1 level by interacting with AP-1 and AP-2.

Identification of LZTFL1 as a candidate effector gene at a COVID-19 risk locus

“…The association of 3p21.31 variants with susceptibility to SARS-CoV-2 infection, as well as disease severity, highlights the importance of the respiratory epithelium for this locus 11 . LZTFL1 encodes a cytosolic leucine zipper protein, which associates with the epithelial marker E-cadherin and is involved in the trafficking of numerous signaling molecules 45,[71][72][73][74] . We note that upregulation of LZTFL1 in the context of malignancy inhibits EMT 45,59,60 , a pathway known to be part of both wound healing and immune responses [53][54][55][56] .…”

Semantic and population analysis of the genetic targets related to COVID-19 and its association with genes and diseases

“…The LZTFL1 gene codes for a leucine zipper protein, which associates with E-cadherin and participates in the circulation of a variety of signaling molecules (84, 85). The gene is expressed in pulmonary epithelial cells, among others, and has been recently identified as a target for a probable causative variant related to COVID-19 risk (86, 87).…”

Effect of CRISPR/Cas9 Targets Associated with Iron Metabolism and Its Variation on Transcriptional Regulation of SHK-1 Cell Line as a Model for Iron Metabolism

“…The AP-1 complex participates in protein-mediated trafficking and has been previously associated with a role in inflammation and gut immune homeostasis [41]. Even more, the AP-1 complex is involved in transferrin receptor 1 trafficking, with direct involvement in the transferrin uptake into the cell [42]. This evidence suggests that different target guides for transferrin could generate not only a general variability on transcriptional regulation but also a specific regulation of processes directly involved in transferrin capture and recognition through affecting membrane trafficking.…”