The Dual-Specificity Kinase DYRK1A Modulates the Levels of Cyclin L2 To Control HIV Replication in Macrophages.

Javan K Kisaka,Lee Ratner,George B Kyei,Guido Silvestri

doi:10.1128/jvi.01583-19

Javan K Kisaka, Lee Ratner + Show 2 more

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https://doi.org/10.1128/jvi.01583-19

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Abstract

HIV replication in macrophages contributes to the latent viral reservoirs, which are considered the main barrier to HIV eradication. Few cellular factors that facilitate HIV replication in latently infected cells are known. We previously identified cyclin L2 as a critical factor required by HIV-1 and found that depletion of cyclin L2 attenuates HIV-1 replication in macrophages. Here we demonstrate that cyclin L2 promotes HIV-1 replication through interactions with the dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A). Cyclin L2 and DYRK1A were colocalized in the nucleus and were found together in immunoprecipitation experiments. Knockdown or inhibition of DYRK1A increased HIV-1 replication in macrophages, while depletion of cyclin L2 decreased HIV-1 replication. Furthermore, depletion of DYRK1A increased expression levels of cyclin L2. DYRK1A is a proline-directed kinase that phosphorylates cyclin L2 at serine residues. Mutations of cyclin L2 at serine residues preceding proline significantly stabilized cyclin L2 and increased HIV-1 replication in macrophages. Thus, we propose that DYRK1A controls cyclin L2 expression, leading to restriction of HIV replication in macrophages.IMPORTANCE HIV continues to be a major public health problem worldwide, with over 36 million people living with the virus. Although antiretroviral therapy (ART) can control the virus, it does not provide cure. The virus hides in the genomes of long-lived cells, such as resting CD4+ T cells and differentiated macrophages. To get a cure for HIV, it is important to identify and characterize the cellular factors that control HIV multiplication in these reservoir cells. Previous work showed that cyclin L2 is required for HIV replication in macrophages. However, how cyclin L2 is regulated in macrophages is unknown. Here we show that the protein DYRK1A interacts with and phosphorylates cyclin L2. Phosphorylation makes cyclin L2 amenable to cellular degradation, leading to restriction of HIV replication in macrophages.

Highlights

human immunodeficiency virus (HIV) replication in macrophages contributes to the latent viral reservoirs, which are considered the main barrier to HIV eradication
To find which domain is critical for interacting with dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A), we constructed three Myc-tagged clones consisting of the full-length cyclin L2, cyclin L2ΔRS, and cyclin L2Δc-box
The data point to a mechanism whereby inhibition of DYRK1A results in increased amounts and a more stable cyclin L2 which promote HIV replication in macrophages

Summary

Introduction

HIV replication in macrophages contributes to the latent viral reservoirs, which are considered the main barrier to HIV eradication. We previously identified cyclin L2 as a critical factor required by HIV-1 and found that depletion of cyclin L2 attenuates HIV-1 replication in macrophages. To get a cure for HIV, it is important to identify and characterize the cellular factors that control HIV multiplication in these reservoir cells. They are synthesized during interphase and rapidly degraded at each mitotic phase [21, 22] They bind to and activate specific CDKs which, in turn, control cell division and transcription [23,24,25]. Using a yeast two-hybrid screen, we previously identified cyclin L2 as a critical factor required for HIV replication in noncycling cells, such as differentiated THP-1 cells and monocyte-derived macrophages (MDMs), but not in dividing cells [34]. It enables the cyclins to localize to nuclear speckles [37, 38], where they are thought to be involved in pre-mRNA processing [33, 37, 39]

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