Cell death may be triggered by ‘hit-and-run’ interaction

A ‘hit-and-run’ interaction between two proteins could be an important trigger for cell death, according to
new research from Walter and Eliza Hall Institute researchers.

The researchers investigated the chain of events that activates the protein BAX, which is a crucial driver
of apoptosis, the major form of cell death. Addressing a long-standing question in the field, they
discovered that BAX is activated for cell death by transient interactions with so-called ‘BH3-only’ proteins
at two distant sites on BAX.

The research, led by Dr Michael Dengler and Professor Jerry Adams, was published today in the journal
Cell Reports.

At a glance

• Activation of the protein BAX is a key step in apoptosis, the programmed death of cells.
• Our researchers revealed a previously controversial ‘hit-and-run’ interaction between BAX and
  ‘BH3-only’ proteins that initiates the conversion of BAX into a lethal protein for cells.
• The discovery, which clarifies a long-standing question in apoptosis research, could underpin the
  development of new approaches to trigger or prevent the death of cells in certain diseases.

Triggering cell death

Apoptosis is the major way our bodies remove damaged or unwanted cells. Many different stimuli trigger
apoptosis by turning on cell signalling pathways that activate BAX and its close relative BAK. Activated
BAX and BAK create holes in the cell’s energy factories, the mitochondria. Once mitochondria are
damaged, cells are compelled to die.

Professor Adams said a long-standing question in apoptosis research had been how BAX is triggered to
move to mitochondria once cell death is triggered.
“The events activating BAX once it has embedded on the surface membrane of mitochondria have been
well-characterised – we know that death-inducing BH3-only proteins bind to BAX, changing its shape to
damage the mitochondrial membrane,” he said.

“There had been hints that BH3-only proteins are also the signal for BAX to move from its location in a
healthy cell’s cytosol – the liquid interior of the cell – to the mitochondria, but the experimental data
supporting this were controversial and weak.”

Two-step activation

To understand how BAX interacts with BH3-only proteins, Dr Dengler and colleagues strategically
altered different regions of BAX, subtly changing the protein’s structure. By comparing the behaviour of
these mutant forms of BAX with that of normal, unmutated BAX, they could determine the function of
different regions of BAX, he said.

“We discovered that two different parts of BAX could bind to BH3-only proteins,” he said. “Intriguingly,
these sites functioned at different stages of BAX activation.

“One site prompted BAX to move to the mitochondrial membrane. Binding of BH3-only proteins to this
site on BAX changed BAX’s structure, releasing a ‘tail’ that anchors BAX to mitochondria. When BH3-
only proteins bound the other site on BAX, BAX became able to damage the mitochondria.”

These two distinct steps in BAX activation had not previously been clearly distinguished.

“The first, early activation step had never been well characterised, because it appears to involve a
transient ‘hit-and-run’ interaction between BAX and a BH3-only protein. We think this first step might be
a way that BAX activation can be fine-tuned,” Dr Dengler said.

The research involved collaborations with structural biology and proteomics researchers at the Institute,
aided by the Australian Synchrotron and the CSIRO Collaborative Crystallisation Centre. “Structural
biology and proteomics were critical technologies for understanding how BAX is activated,” Dr Dengler
said.

As well as explaining a key detail in how cell death is executed, the research may in the future lead to
new classes of drugs that modify BAX function.

“BAX is a key mediator of cell death, and many major diseases involve either too little or too much cell
death. Our discovery may eventually underpin the search for drugs that promote apoptosis by activating
BAX, which may have potential for treating cancer. Conversely, drugs that block BAX activation could
help to prevent the harmful cell death that occurs in neurodegenerative disorders or stroke,” Professor
Adams said.

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The research was supported by the Australian National Health and Medical Research Council, the USbased
Leukemia and Lymphoma Society and the Victorian Government.

For more information contact the Institute media office on 0475 751 811 or [email protected]