ZNF330/NOA36 interacts with HSPA1 and HSPA8 and modulates cell cycle and proliferation in response to heat shock in HEK293 cells.

BACKGROUND: The human genome contains nearly 20.000 protein-coding genes, but there are still more than 6,000 proteins poorly characterized. Among them, ZNF330/NOA36 stand out because it is a highly evolutionarily conserved nucleolar zinc-finger protein found in the genome of ancient animal phyla like sponges or cnidarians, up to humans. Firstly described as a human autoantigen, NOA36 is expressed in all tissues and human cell lines, and it has been related to apoptosis in human cells as well as in muscle morphogenesis and hematopoiesis in Drosophila. Nevertheless, further research is required to better understand the roles of this highly conserved protein. RESULTS: Here, we have investigated possible interactors of human ZNF330/NOA36 through affinity-purification mass spectrometry (AP-MS). Among them, NOA36 interaction with HSPA1 and HSPA8 heat shock proteins was disclosed and further validated by co-immunoprecipitation. Also, "Enhancer of Rudimentary Homolog" (ERH), a protein involved in cell cycle regulation, was detected in the AP-MS approach. Furthermore, we developed a NOA36 knockout cell line using CRISPR/Cas9n in HEK293, and we found that the cell cycle profile was modified, and proliferation decreased after heat shock in the knocked-out cells. These differences were not due to a different expression of the HSPs genes detected in the AP-MS after inducing stress. CONCLUSIONS: Our results indicate that NOA36 is necessary for proliferation recovery in response to thermal stress to achieve a regular cell cycle profile, likely by interaction with HSPA1 and HSPA8. Further studies would be required to disclose the relevance of NOA36-EHR interaction in this context.

Zinc finger protein 330 regulates Ramie mosaic virus infection in the whitefly Bemisia tabaci MED.

BACKGROUND: The whitefly, Bemisia tabaci (Gennadius) is one of the most economically important pests that cause serious damage to agricultural production by transmitting plant pathogenic viruses. Approximately 90% of the virus species transmitted by the whitefly are members of the genus begomovirus. Ramie mosaic virus (RaMoV) is a new bipartite begomovirus that causes severe damage to ramie and several other economic crops in China. In previous studies, we have demonstrated that RaMoV had no obvious direct or indirect effects on B. tabaci. However, whether B. tabaci affects RaMoV infection and the molecular mechanisms of their interaction remain unclear. RESULTS: Here, we identified a zinc finger protein 330 (ZNF330) in B. tabaci MED interacted with the coat protein (CP) of RaMoV by the yeast two-hybrid assay. Then the interaction between ZNF330 and RaMoV CP was further verified by glutathione S-transferase (GST) pull-down assay. The expression of ZNF330 gene was continuously induced after RaMoV infection. ZNF330 negatively regulated RaMoV replication in the B. tabaci MED. Furthermore, the longevity and fecundity of RaMoV-infected female adults were significantly decreased after silencing of ZNF330. CONCLUSIONS: Our results indicated that the ZNF330 protein was involved in the negative regulation of RaMoV replication in the B. tabaci MED. High viral accumulation caused by ZNF330 silencing is detrimental to fecundity and longevity of the B. tabaci MED. These findings provided a new insight into identifying the binding partners in whitefly with viral CP and fully understanding the complex interactions between begomoviruses and their whitefly vector. © 2023 Society of Chemical Industry.

ZNF330 promotes erythroid differentiation / 基础医学与临床

Objective To explore the effects of ZNF330 on erythroid differentiation of K562 cells and underlying mechanism .Methods Realtime PCR was performed to detect the expression of ZNF 330 in K562 cells induced by hemin .After CD34 +cells being infected by the recombination lentivirus ZNF 330-RNAi, Realtime PCR was applied to detect the expression of CD235a and γ-globin.The luciferase report assay was performed to examine if ZNF 330 could act as a trans-acting factor in 293T/17 cells.Co-Immunoprecipitation (Co-IP) was applied in 293T/17 cells to detect the interaction between ZNF 330 and ZNF408 which was involved in mRNA degradation .Results The ex-pression of ZNF330 was up-regulated after hemin treatment .The expression of CD235a andγ-globin decreased after inhibition expression of ZNF 330 had no effect on report gene .Co-Ip in two ways confirmed the direct binding be-tween ZNF330 and ZNF408 .Conclusions ZNF330 can promote erythroid differentiation , and a possible mecha-nism is that ZNF330 inhibits the function of ZNF 408 , a factor that is involved in mRNA degradation , through the interaction between the two proteins .