Zinc finger protein 64 promotes Toll-like receptor-triggered proinflammatory and type I interferon production in macrophages by enhancing p65 subunit activation
Authors: Peng Zhang
Xingguang Liu
Zhenyu Yao
Qian Zhang
Nan Li
Bo Huang
Yin Liu
Chunmei Wang
Xuetao Cao
Journal: Journal of Biological Chemistry (2013)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Inflammation; Innate Immunity; Macrophages; NF-κB (NF-κB); Toll-like Receptors (TLR)
Affiliations: National Key Laboratory of Medical Molecular Biology and Department of Immunology, Chinese Academy of Medical Sciences, Beijing 100005, China.

Abstract:  The molecular mechanisms that fine-tune the Toll-like receptor (TLR)-triggered innate immune response need further investigation. As an important transcription factor, zinc finger proteins (ZFPs) play important roles in many cell functions, including development, differentiation, tumorigenesis, and functions of the immune system. However, the role of ZFP members in the innate immune responses remains unclear. Here we showed that the expression of C2H2-type ZFP, ZFP64, was significantly up-regulated in macrophages upon stimulation with TLR ligands, including LPS, CpG oligodeoxynucleotides, or poly(I:C). ZFP64 overexpression promoted TLR-triggered TNF-α, IL-6, and IFN-β production in macrophages. Coincidently, knockdown of ZFP64 expression significantly inhibited the production of the above cytokines. However, activation of MAPK and IRF3 was not responsible for the ZFP64-mediated promotion of cytokine production. Interestingly, ZFP64 significantly up-regulated TLR-induced NF-κB activation. ZFP64 could bind to the promoter of the TNF-α, IL-6, and IFN-β genes in macrophages only after TLR ligation. Furthermore, ZFP64 associated with the NF-κB p65 subunit upon LPS stimulation, and TLR-ligated macrophages showed a lower level of p65 recruitment to the TNF-α, IL-6, and IFN-β gene promoter in the absence of ZFP64. The data identify ZFP64 as a downstream positive regulator of TLR-initiated innate immune responses by associating with the NF-κB p65 subunit, enhancing p65 recruitment to the target gene promoters and increasing p65 activation and, thus, leading to the promotion of TLR-triggered proinflammatory cytokine and type I interferon production. Our findings add mechanistic insight into the efficient activation of the TLR innate response against invading pathogens. 

Retracted: No

PubPeer comments:
 Posting for someone else. A reader pointed out an unexpected similarity between the actin panel in Figure 2A and the first two lanes of the actin panel in Figure 4B. These appear to represent different experiments. Shown with red boxes. ![file] , 
 Figure 1A appears to show unexpected similarities between some lanes in the ZFP64 panel. In addition, all lanes in the beta-actin panel appear to show similar dots above and below the bands. Could the authors please check what might have been causing this? (This is my own question, not part of what the reader mentioned in #1 sent) ![file] , 
 2020 Expression of Concern. https: www.jbc.org content 295 26 8885 The publisher of the Journal of Biological Chemistry is issuing an Expression of Concern to inform readers that credible concerns have been raised regarding some of the data and conclusions in the article listed above. The Journal of Biological Chemistry will provide additional information as it becomes available. , 
 Retraction on Jan 1, 2021, published online May 22, 2021 https: www.jbc.org article S0021-9258(21)00549-4 fulltext Withdrawal: Zinc finger protein 64 promotes toll-like receptor-triggered proinflammatory and type I interferon production in macrophages by enhancing p65 subunit activation This article has been withdrawn by the authors. The Journal states that in Fig. 1B lanes 2 and 3 from both panels were identical. Lanes 4 and 5 in Fig. 1B, lower panel, were also duplicated. Lanes 5 and 6 in Fig. 1B, upper panel, were duplicated. The actin immunoblot in the first two lanes of Fig. 4B was reused in Fig. 2A, representing different experimental conditions. In Fig. S5, the first four lanes of the HA immunoblot panel (right) were reused as the last four lanes of the Flag panel immunoblot (right). For Fig. 1B, Fig. 4B-2A and Fig. S5, the authors are able to retrieve the raw data and offer to update the respected panels using the same raw images again or those from another experiment with the same condition. The offer was declined by the Journal. The authors state that the issues described above do not affect the interpretation of the data nor conclusions of this research study. The authors have indicated that they plan to submit this work elsewhere for publication. ,